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Silk fibroin/stem cell compounded microsphere and application thereof

A technology of silk fibroin microspheres and composite microspheres is applied in the application field of silk fibroin/stem cell composite microspheres and as injection fillers, which can solve the problem that polymer synthetic materials do not have biocompatibility and lack mechanical and mechanical properties. It can improve structural and functional defects, lasting therapeutic effects, and promote local muscle and nerve regeneration.

Inactive Publication Date: 2014-05-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mechanical properties of collagen are not good, and polymer synthetic materials do not have good biocompatibility

Method used

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  • Silk fibroin/stem cell compounded microsphere and application thereof
  • Silk fibroin/stem cell compounded microsphere and application thereof
  • Silk fibroin/stem cell compounded microsphere and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 silk fibroin microsphere

[0044] 1) Preparation of silk fibroin solution:

[0045] a) Degumming: put silkworm silk (Zhejiang Huazhi Silk Co., Ltd.) into 2M aqueous sodium carbonate solution, bathe in 98°C water for 30 minutes, and wash with purified water. Repeat this process 3 times to remove sericin and leave silk fibroin Protein fibers, drying silk fibroin fibers at 50°C for later use;

[0046] b) Dissolution: Dissolve the above-mentioned dried silk fibroin in 9.3M lithium bromide (LiBr) aqueous solution at a mass volume concentration of 20%, and bathe in water at 60°C for 120 minutes until the silk fibroin is fully dissolved;

[0047] c) Dialysis: Dialyze the mixed solution with a regenerated cellulose dialysis bag (molecular weight cut-off 8000 Daltons), and use purified water 10 times the volume of the silk solution for 10-12 times in 3 days to remove LiBr ions in the solution;

[0048] d) Centrifugation: centrifuge in a horizontal rotor at 5,000g ...

Embodiment 2

[0056] Pass c) in step 2) of Example 1 through an 80-mesh sieve to make silk fibroin microspheres with a particle size of 180 μm, and prepare a suspension of 2 g / ml silk fibroin microspheres and water.

Embodiment 3

[0057] Example 3 Isolation, cultivation and identification of adipose-derived stem cells

[0058] Reference: Shi L B, Cai H X, Chen L K, et al.Tissue engineered bulking agent with adipose-derived stem cells and silk fibroin microspheres for the treatment of intrinsic urethral sphincter deficiency[J].Biomaterials,2014,35(5):1519 -1530.

[0059] 1) Isolation and culture of adipose-derived stem cells

[0060] a) 4-week-old female SD rats were anesthetized by intraperitoneal injection of 4% chloral hydrate 4ml / kg, skin was prepared, sterilized by soaking in 75% ethanol for 3min, the inguinal fat pad was cut out, and PBS buffer (pH 7.0) wash 3 times;

[0061] b) Fully shred the fat pad, add 3 times the volume of 0.25% mixed collagenase (Sigma-Aldrich Inc., St. Louis, MO, USA), shake and digest at 37°C for 30 minutes;

[0062] c) Dilute 5 times with DMEM low-sugar medium containing 10% FBS and 1% PS by volume, centrifuge at 1000r / min for 5min, discard the upper layer, remove the ...

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Abstract

The invention discloses a silk fibroin / stem cell compounded microsphere and application thereof. The silk fibroin / stem cell compounded microsphere is prepared with the following method: silk fibroin microsphere suspension and stem cell suspension are sufficiently mixed, and then placed in a sterile culture dish and cultured in a cell incubator in conditions that the temperature is 37 DEG C, CO2 content is 5 percent, and the saturated humidity is 100 percent, a culture solution is changed every 3 days, and culture is performed till silk fibroin microsphere and stem cells are compounded to form a cell microsphere. Urethra tissue engineering filler constructed through an ADSCs composite silk fibroin microsphere is used for curing SUI rat, on one hand, by injecting the silk fibroin microsphere, supporting and filling are provided for vesical necks, and an instant treatment effect is produced; on the other hand, regeneration of local muscle and nerves is promoted and the structural and functional defects of urethral sphincter are improved through the transplantation of ADSCs compounded on the silk fibroin microsphere and subsequent cell differentiation and multiplication to improve the urine control ability of urethral sphincter, so that the lasting treatment effect is achieved.

Description

(1) Technical field [0001] The invention relates to a tissue engineering injection filler, in particular to a silk fibroin / stem cell composite microsphere and its application as an injection filler, which can be used for minimally invasive treatment of stress urinary incontinence. (2) Background technology [0002] Stress urinary incontinence (SUI) refers to the involuntary leakage of urine from the external opening of the urethra when abdominal pressure increases, such as sneezing or coughing. According to the epidemiological survey results of various countries, the prevalence rate of women aged 15-64 in the United States is 2%-46%, and the prevalence rate of women over 60 years old in the United Kingdom is 69%. According to the latest survey statistics in my country, the incidence rate of SUI in women is as high as 22.9%, while middle-aged and elderly women over 40 years old, the incidence of SUI has risen to 35.8%, and the incidence is still increasing rapidly with the agi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/22A61L27/50
Inventor 邹晓晖施丽冰
Owner ZHEJIANG UNIV
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