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A kind of composite support material

A technology of composite scaffolds and composites, applied in the field of tissue engineering, can solve problems such as low content, limited source of stem cells, and large body trauma

Active Publication Date: 2017-09-08
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the source of bone marrow mesenchymal stem cells is limited and the content is low. The extraction of bone marrow requires bone puncture to obtain it, which causes great trauma to the body. Moreover, the source of bone marrow is limited, which contains complex cellular components, and the content of stem cells is low. Requires a longer period of expansion and culture in vitro
Therefore, the scope of clinical application of bone marrow mesenchymal stem cells is relatively narrow
At the same time, this study found that bone marrow mesenchymal stem cells have low osteogenic differentiation ability, poor composite ability with scaffold materials, and difficulty in mineralizing nodules, which will further affect the effect of bone defect treatment

Method used

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  • A kind of composite support material
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 The acquisition of dental pulp stem cells

[0051] With the consent of the family members, clinically collected deciduous teeth extracted from children due to detention, and no dental pulp disease and necrosis were required. Immediately after pulling out, put it in a PBS centrifuge tube containing double antibody when it is cold at 4°C, and take it out for use within 24 hours. The teeth were taken out, rinsed three times repeatedly with PBS solution containing double antibodies, wrapped in sterile gauze and cracked with forceps; the pulp tissue was clamped with sterile tweezers, and 1mm of the pulp tissue at the apex was excised; Curved scissors cut the pulp tissue into 1mm 3 , put in a 50mL centrifuge tube, add 10 times the volume of 5g / L type I collagenase, seal the seal and mix well, transfer to a constant temperature air shaker, digest at 37°C and 200R for 10-20min; add an equal volume of serum-free culture Repeatedly blow and beat the discrete cell aggr...

Embodiment 2

[0053] Example 2 Acquisition of bone marrow stem cells

[0054] With the consent of the family members and the person, the doctor uses a bone marrow puncture trocar to aspirate 5-8 mL of bone marrow, put it in an anticoagulant tube containing serum-free medium, rotate it gently and mix well, and then take it back to the laboratory for processing; In the ultra-clean workbench, transfer the uncoagulated bone marrow into a centrifuge tube, add the same volume of serum-free medium, centrifuge at 1000rpm for 20min; discard the supernatant and tissue fluid, resuspend in 2mL of serum-free medium, and transfer it into a new sterile tube. Mix well in a centrifuge tube, centrifuge at 1000rpm for 10min; discard the supernatant, add 15% FBS DMEM / F12 medium equal to the volume of the precipitate to resuspend, dilute 30-50 times, inoculate in a petri dish for culture, and store at 37°C and humidity Cultivate in a 95% carbon dioxide incubator; change the medium every day for the first three ...

Embodiment 3

[0056] Example 3 In vitro osteogenesis induction experiment

[0057] The dental pulp mesenchymal stem cells obtained in Example 1 and the bone marrow mesenchymal stem cells obtained in Example 2 that grew well and approached confluence in the P3 generation were respectively taken, digested with trypsin, and made a cell suspension with a new medium, and then counted. Adjust the cell density to 1 x 10 5 / mL, according to 1×10 4 cell / cm 2 Seed into 6-well plate, 2mL / well. When the cells grew to 80% confluence, the culture medium was discarded and replaced with osteogenic induction medium, 2 mL / well (the control group was cells, and ordinary medium was used instead of induction factor medium). Continuous induction, the induction medium was replaced every 3 days, and the induction was stopped on the 28th day, and the cell changes were detected by Alizarin red staining and the dentin matrix protein 1 (DMP1) and sclerostin (SOST) were detected by Real-time fluorescent quantitative...

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Abstract

The invention relates to the technical field of tissue engineering, and in particular relates to a composite stent material. A preparation method of the composite stent material comprises the steps that a chitosan solution is mixed with an I-type collagen solution; pH is kept acidic; a chitosan / I-type collagen compound is obtained, mixed with a cross-linking agent, and subjected to freeze drying; a multi-hole stent is obtained, and soaked in a serum-free medium; the serum-free medium is removed; a dental pulp stem cell suspension liquid is added; standing is performed; a complete medium is added for cell culture; and the composite stent material is obtained. The research of composite stent material shows that a dental pulp stem cell has better osteogenic differentiation capacity than a bone marrow stem cell; and the dental pulp stem cell can be tightly combined with the stent material, grows well in the stent material well, and can be applied to treatment on a bone defect better.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a composite bracket material. Background technique [0002] Bone defect refers to a disease in which the integrity of bone structure is destroyed due to trauma, infection, tumor, surgical debridement of osteomyelitis, and various congenital diseases. It is a common and frequently-occurring disease in clinical practice. The treatment of bone defects is mainly through autologous bone grafting, allogeneic bone grafting, artificial bone grafting, tissue engineering bone grafting, bone handling technology, etc. Among them, transplantation using autologous bone and artificial bone has its own shortcomings. The source of autologous bone is limited, the extra blood loss during the operation and the time of surgical anesthesia are longer, and complications such as donor site fracture and bone pain may occur; although the source of allogeneic bone or artificial bone is relative...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/26A61L27/24A61L27/20A61L27/38A61L27/56
Inventor 葛啸虎陈海佳王一飞马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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