Decellularized fiber ring matrix preparation method

Abstract

The invention discloses a decellularized fiber ring matrix preparation method and belongs to the technical field of animal cells or tissues. The decellularized fiber ring matrix preparation method includes taking spinal fiber ring of a healthy adult pig, sufficiently rinsing the spinal fiber ring by sterile saline solution; placing the spinal fiber ring into Tris buffer solution of concentration 10mM and with 0.05-0.5% of EDTA (ethylene diamine tetraacetic acid) and 5-50KIU/ml of aprotinin and oscillating at the temperature of 4 DEG C; then placing the spinal fiber ring into the Tris buffer solution containing 1-5% of TritonX-100, oscillating at room temperature; placing the spinal fiber ring into Tris buffer solution containing 0.05-0.5mg/ml of deoxyribonuclease, 5-50microgram/ml of ribonuclease A, and oscillating; and finally, washing by sterile saline solution with oscillating, and storing in sterile PBS (phosphate buffer solution) at the temperature of 4 DEG C. The decellurlarized fiber ring matrix preparation method is simple in preparation process, thorough in decellurlarization and complete in reserved structure, similar in matrix content and mechanical performance as those of natural fibers, and is a good tissue engineering fiber ring support material.

Description

technical field [0001] The invention relates to the field of animal cells or tissues, in particular to a method for preparing an acellular fibrous annulus matrix. Background technique [0002] Intervertebral disc degeneration is a common disease that seriously affects people's lives, and it is also one of the difficult problems in orthopedic treatment. The emergence of tissue engineering technology provides new ideas and methods for the treatment of intervertebral disc diseases, and is expected to provide permanent replacement products for intervertebral disc repair or replacement. Among them, the regeneration and repair of the annulus fibrosus is the first problem to be solved in intervertebral disc tissue engineering, and only the annulus fibrosus can be obtained. Repair can keep the nucleus pulposus tissue in the intervertebral space, and then carry out the next step of treatment for the degenerated nucleus pulposus. [0003] In recent years, there have been many studies...

AI Technical Summary

Problems solved by technology

For example, oriented electrospun nanoscaffolds have regular structure and uniform pores, but their composition is quite different from that of natural fibrous annulus, and their biocompatibility is poor; the mechanical properties of oriented alginate / chitosan composite scaffolds are similar to those of natural fibrous annulus. The difference is far; polycaprolactone malate triol / decalcified bone matrix gelatin biphasic fibrous annular scaffold, the inner and outer layers of the biphasic scaffold are not firmly bonded, easy to fall off, and have poor biocompatibility; domestic Pan Yong et al. Mineral decellularized bone matrix ring is used as the fibrous ring scaffold, which has good cytocompatibility, but its plasticity is poor, and its structure is far from the complex structure of the natural fibrous ring.

In summary, the biochemical composition and structure of the extracellular matrix (ECM) of the annulus fibrosus are complex and arranged in oblique layers. However, at present, it is difficult to truly imitate the complex natural structure, composition and biological characteristics of the intervertebral disc, no matter artificially synthesized or natural biomimetic materials at home and abroad. Mechanical properties

There is no report on the use of acellular annulus fibrosus matrix as an annulus fibrosus scaffold

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