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Technical means for thermo-responsive hydrogel package delivery of exosomes and treatment effect enhancement

A hydrogel and responsive technology, applied in liquid delivery, ointment delivery, drug delivery, etc., can solve the problems of unclear stability of bioactive molecules and low retention rate of exosomes, and achieve enhanced retention rate and stability Sexuality, the effect of improving the therapeutic effect

Inactive Publication Date: 2018-11-06
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in practical application, due to the small diameter of exosomes, they are easily swallowed by the immune system and the reticuloendothelial system, and the retention rate of exosomes at the injury site after injection is very low; at the same time, although they are stored at low temperature, The components in exosomes are very stable, but the stability of the bioactive molecules carried by exosomes after they are injected into the body is still unclear

Method used

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  • Technical means for thermo-responsive hydrogel package delivery of exosomes and treatment effect enhancement
  • Technical means for thermo-responsive hydrogel package delivery of exosomes and treatment effect enhancement
  • Technical means for thermo-responsive hydrogel package delivery of exosomes and treatment effect enhancement

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Embodiment 1

[0019] Embodiment 1, the present invention provides a method for extraction and identification of stem cell-derived exosomes.

[0020] Cell culture medium, double antibody, trypsin and other reagents were purchased from Gibco Company; cell culture consumables were purchased from Corning Company.

[0021] The serum required for cell culture is fetal bovine serum with exosomes removed, purchased from BI company, and the processing steps are as follows: put the fetal bovine serum in an ultracentrifuge tube, centrifuge at 120,000 g at 4 °C for 2 h, and take it in an ultra-clean bench. and filtered with a 0.22 μm syringe filter, and stored in a -80°C refrigerator for later use.

[0022] For cell biology experiments such as cell subculture, cryopreservation and recovery, please refer to "Animal Cell Culture (Sixth Edition)".

[0023] Collect hP-MSCs conditioned medium containing exosomes: When hP-MSCs cultured in a 75 cm2 cell culture flask is in the logarithmic growth phase and th...

Embodiment 2

[0035] Embodiment 2, the present invention provides a preparation method of a temperature-responsive hydrogel.

[0036] Preparation method of temperature-responsive chitosan hydrogel: Dissolve chitosan (Mn = 50,000) powder in 0.1 M acetic acid, filter sterilize with 0.22 μm filter, and prepare a solution of 2% chitosan in an ice bath . Make up a 50% beta-glycerophosphate solution and filter sterilize. In an ice bath, add the β-glycerophosphate solution to the chitosan solution at a volume ratio of 1:5 with constant stirring until the two solutions are well mixed. The chitosan / glycerophosphoric acid mixed solution can be cross-linked into a hydrogel after incubating at 37°C for 30 minutes (such as figure 1 shown in A).

[0037] Preparation method of temperature-responsive collagen fiber gel: take SD rats weighing about 250 g and take their tails, soak them in 75% ethanol for sterilization, break the coccyx and remove the fur under aseptic conditions, extract the tail tendons...

Embodiment 3

[0038] Embodiment 3, the present invention provides a method for encapsulating and delivering exosomes using a temperature-responsive hydrogel.

[0039] Dissolve 2× hydrogel solution and stem cell-derived exosomes in equal volume to prepare 1× hydrogel solution containing exosomes, and keep it at 4°C all the time. Keep the syringe at 4°C during use. ℃ pre-cooling, after injection into the injury site, cross-linking in vivo at 37 ℃ to form a hydrogel containing exosomes.

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Abstract

The invention discloses a technical means for thermo-responsive hydrogel package delivery of exosomes and treatment effect enhancement. The technical means includes: wrapping stem cell derived exosomes with thermo-responsive hydrogel which is liquid at 4 DEG C, injecting to an injured part to form hydrogel with wrapped exosomes in vivo, and slowly releasing the exosomes to injured tissues. By adoption of the hydrogel, the retention rate and local concentration of the exosomes at the injured part can be increased while stability of functional molecular in the exosomes is improved, and the exosomes can be slowly released to the injured part to further improve treatment effects of the exosomes and promote structural and functional recovery of the injured tissues.

Description

technical field [0001] The invention relates to a technical means for delivering exosomes by wrapping a temperature-responsive hydrogel and enhancing their therapeutic effect in tissue damage, and belongs to the technical field of tissue engineering and new medicine. Background technique [0002] Exosomes are the main form of paracrine components and play an important role in intercellular communication. Exosomes are generally considered to be microvesicles produced by the inward budding of multivesicular bodies inside cells. When the multivesicular bodies fuse with the cell membrane, the inner microvesicles are released to the outside of the cell membrane to form exosomes. . The diameter of exosomes is about 30-100 nm. Due to their special formation, exosomes are rich in intracellular biologically active signaling molecules, including DNA, RNA, microRNA, and proteins. The intercellular communication mediated by exosomes can provide a large number of biologically active ma...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K35/34A61K35/44A61K35/22A61K9/06A61P9/10A61P13/12A61P17/02
CPCA61K9/0019A61K9/06A61K35/22A61K35/28A61K35/34A61K35/44A61P9/10A61P13/12A61P17/02
Inventor 李宗金张凯悦
Owner NANKAI UNIV
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