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Method for preparing double-layer scaffold by single-step process and method for tissue regeneration using the double-layer scaffold obtained by the preparation method

A double-layer, polymer technology, used in tissue regeneration, layered products, skin transplantation, etc., can solve the problems of large-scale cell culture, inconvenient preparation methods, and complex processes.

Active Publication Date: 2018-05-29
RES COOPERATION FOUND OF YEUNGNAM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many problems with the above-mentioned scaffolds: its adhesion is not enough for mass culture of cells, its physical properties are not outstanding, and its preparation method is not convenient and satisfactory
[0006] In addition, conventional bilayer scaffolds are prepared by attaching each polymer scaffold to each other after fabricating them separately, however, the bilayer scaffold cannot be completely attached, there is a great possibility of separation, and, due to the The preparation is a two-step method, and the process is very complicated

Method used

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  • Method for preparing double-layer scaffold by single-step process and method for tissue regeneration using the double-layer scaffold obtained by the preparation method
  • Method for preparing double-layer scaffold by single-step process and method for tissue regeneration using the double-layer scaffold obtained by the preparation method
  • Method for preparing double-layer scaffold by single-step process and method for tissue regeneration using the double-layer scaffold obtained by the preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] preparation of double-layer stent

[0060] 1) Preparation of double-layer scaffold

[0061] A 2.5 wt% aqueous solution of gelatin was prepared and stirred at 300 rpm at 80°C. After the gelatin was completely dissolved, polyvinyl alcohol (PVA) was added to make it 10 wt%, and stirred. Sodium lauryl sulfate was added so as to be 0.2 wt%, and stirred at a high speed of 900 rpm at a high temperature of 95°C. The stirred solution was poured into Petri dishes and lyophilized at -80°C for at least 24 hours to obtain sponges. The sponge was crosslinked by soaking it in an aqueous solution containing glutaraldehyde (0.5 wt %) for at least 12 hours. Freeze drying again at -80°C yielded a bilayer scaffold.

[0062] 2) SEM and digital images of bilayer scaffolds

[0063] By means of a field emission scanning electron microscope (FE-SEM) (S-4100, HITACHI, LTD) the double-layer scaffold prepared as above is observed, and the SEM image obtained is shown in figure 2 Middle; dig...

experiment Embodiment 1

[0070] In vitro performance of double-layer stent

[0071] After the double-layer stent prepared in the embodiment was made into a diameter of 1 cm, 2×10 4 CRL-2310 (ATCC number; keratinocytes, human papillomavirus 16 (HPV-16) E6 / E7 transformed) partitioned on the bilayer scaffold. After cell allocation, as a result of SEM observation, such as Figure 7 As shown, cells began to attach to the scaffold and regenerate 1 day after cell dispensation; as Figure 8 As shown, accumulation of extracellular matrix (ECM) secreted by cells into the scaffold was confirmed 5 days after cell allocation.

experiment Embodiment 2

[0072] In vivo wound regeneration effect of double-layer scaffold

[0073]A 7 mm wound was made using a biopsy punch and a bilayer scaffold having the same dimensions as the wound (7 mm) was bent so that the gelatin layer was in contact with the wound site. In this case, the scaffolds were submerged in 20%, 40%, 60%, 80% and 100% ethanol for 1 h for sterilization, and finally in 70% ethanol for 24 h and kept at room temperature. Submerged in PBS buffer for 3 hours before use in experiments. In the control group the wounds were kept open; whereas in the test group the fabricated wounds were covered with the scaffold according to the invention.

[0074] As a result, such as Figure 9 with Figure 10 As shown, from the photographs 1 day after treatment with the double-layer scaffold, the tissue began to bleb from the subcutaneous area, while at least 90% of the wound was maintained in the control group, and the size of the wound was reduced to 80% in the test group. From the...

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Abstract

Disclosed is a method for preparing a bilayer scaffold through single process comprising: preparing a first polymer aqueous solution; adding a second polymer into the first polymer aqueous solution and stirring a reactant; adding a surfactant into the stirred reactant and stirring the reactant at high temperature and high speed; freeze-drying the stirred reactant thereby obtaining a sponge; dipping the sponge in a cross-linking agent thereby rendering be cross-linked; and freeze-drying the cross-linked reactant.

Description

technical field [0001] The present invention relates to a method for preparing a double-layer scaffold through a single-step process and a method for tissue regeneration using the double-layer scaffold obtained by the preparation method. Background technique [0002] Scaffolds are physical supports and attachment substrates prepared for in vitro culture and transplantation of tissue cells; scaffolds are used for cell transplantation for human tissue regeneration; and scaffolds are very important for mass culture and proliferation of cells. The reason for this is that the cell attachment of epithelial cells and the concomitant migration and proliferation occur at the contact portion between the cells and the substrate. [0003] When in contact with substances in vivo or in vitro, most biologically active cells first survive by cell attachment. In particular, in the survival of fibroblasts and tissue cells, cells preferentially attach to the substrate, and then organelles in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/56A61L27/14A61L27/20A61F2/28
CPCA61F2/30756A61F2002/30011A61L27/26A61L27/3687A61L27/3691A61L27/3817A61L27/3821A61L27/56A61L27/60A61L2430/34B32B5/32B32B2250/22C08J9/0061C08J9/28C08J9/30C08J2201/026C08J2201/0484C08J2207/10C08J2301/00C08J2305/00C08J2323/08C08J2325/06C08J2329/02C08J2329/04C08J2333/02C08J2339/06C08J2367/04C08J2371/02C08J2375/06C08J2379/02C08J2389/00C08L89/06C08L29/04A61F2/105A61F2/28A61F2210/0004A61F2210/0076A61F2240/001A61L27/3813A61L2300/412C08J3/24C08J9/36C08J2489/00
Inventor 韩星洙崔顺模迪普·蒂·辛格
Owner RES COOPERATION FOUND OF YEUNGNAM UNIV
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