Unlock instant, AI-driven research and patent intelligence for your innovation.

Acellular ligament stent and seed cell compound culture method

A technology of seed cells and culture methods, which is applied in the field of combined culture of decellularized ligament scaffolds and seed cells, which can solve the problems of small amount, limited application of ligament decellularized scaffolds, and limitations

Active Publication Date: 2013-04-03
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(see literature 1. GRATZER PF, HARRISON RD, WOODS T. Matrix alteration and not residual sodium sulfate cytotoxicity affects the cellular repopulation of a decellularized matrix. Tissue-Eng. 2006; 12 (10): 2975-83.2. Brown AL, Brook-Allred TT, Waddell JE, White J, Werkmeister JA, Ramshaw JA, Bagli DJ, Woodhouse KA. Bladder cellular matrix as a substrate for studying in vitro bladder smoothmuscle-urothelial cell interactions. Biomaterials. 2005; 26(5): 529-43. ) using ultrasound (see figure 2 ), multipoint injection (see image 3 ), oxidation and other methods to increase the porosity of the ligament or tendon decellularized scaffold, but the seed cells in the scaffold are unevenly distributed, less and limited
This limits the application of ligament decellularized scaffolds

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Acellular ligament stent and seed cell compound culture method
  • Acellular ligament stent and seed cell compound culture method
  • Acellular ligament stent and seed cell compound culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0022] Example: Compound culture method and evaluation of rabbit tissue engineering acellular ligament scaffold and seed cells

[0023] 1. Materials: select New Zealand rabbits (about 3kg, provided by the Animal Experiment Center of the Second Military Medical University, the same below), completely resect the patellar ligament and its tibiofibular insertion, remove the outer synovial tissue of the ligament, and ensure the integrity of the outer membrane of the ligament. Store in sterile PBS at 4°C.

[0024] 2. Decellularization

[0025] (1) 10mM hypotonic tris buffer + serine protease inhibitor (0.05% phenylmethylsulfonyl fluoride ethanol solvent, 35ml / L 50ml ethanol + 25mgPMSF) for 48h.

[0026] (2) 0.01% trypsin + 0.02% EDTA 8h

[0027] (3) 1% sodium deoxycholate (DCA) 24h

[0028] (4) Hank's buffer + nuclease (DNase I 200μg / ml+RNase I 20μg / ml) for 5h

[0029] (5) Store at -80°C after washing with PBS for 48 hours

[0030] All steps were carried out on a shaker (150RPM...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of tissue engineering, in particular to an acellular ligament stent and a seed cell compound culture method. The main reasons that the application of an acellular ligament stent in the ligament tissue engineering is limited are that the ligament acellular stent has small porosity and fibroblasts cultured in vitro cannot grow toward the inside of the stent. In the method of the invention, a gap among collagenous fibers can be properly enlarged easily by physical methods such as extrusion and the like after acellular ligaments are dried at the low temperature and the dried acellular ligaments are convenient for seed cell suspension to permeate so as to directly enable seed cells to enter the acellular ligament stent by a permeating method. By culturing in vitro for one week, the cells after HE staining are attached to the collagenous fiber to grow inside the acellular ligaments. The method of the invention is beneficial for the seed cells to enter the ligament stent along with permeation of the cell suspension to grow.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a compound culture method of decellularized ligament scaffold and seed cells. Background technique [0002] Poor healing after repair after ligament and tendon injury is a common clinical problem, which has led to the application of various substitutes for ligament and tendon repair and reconstruction. Among them, allogeneic, heterogeneous ligament and tendon graft material scaffolds have the inherent characteristics of ligament and tendon scaffolds. With the development of decellularization technology and sterilization technology, the immune rejection and disease infection of hosts to allogeneic and heterogeneous ligament scaffolds have been greatly reduced, making ligaments And the clinical application of tendon decellularized scaffolds becomes possible. The current methods for preparing decellularized ligament scaffolds from ligaments and tendons often combine phys...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36C12N5/071
Inventor 陈雄生朱巍周盛源
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY