Nucleus pulposus-cartilage extracellular matrix scaffold and preparing method thereof

A chondrocyte and extracellular matrix technology, which is applied in medical science, prosthesis, etc., can solve the problems such as the mixed preparation of scaffolds without acellular cartilage matrix, the difficult storage of nucleus pulposus extracellular matrix suspension, and the long acellular cycle and easy deterioration. Achieve the effect of simple preparation process, good clinical application prospect, good biocompatibility and degradability

Inactive Publication Date: 2015-11-11
TIANJIN HOSPITAL
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

In a patent (Application No.: 201310000722.2) of "annulus fibrosus and nucleus pulposus integrated two-way scaffold and its construction method", another scholar used the decellularized nucleus pulposus matrix as the nucleus pulposus phase to prepare a new type of integrated annulus fibrosus-nucleus pulposus A biphasic scaffold, but the preparation method of the decellularized nucleus pulposus matrix has the disadvantages that decellularization is not easy to complete, the decellularization cycle is long and easy to deteriorate, and the obtained nucleus pulposus extracellular matrix suspension is not easy to store and the concentration is not easy to adjust.
Moreover, the integrated annulus fibrosus-nucleus pulposus biphasic scaffold is suitable for the repair of total intervertebral disc replacement, while clinically, multiple intervertebral disc partial defects
At present, there is no report on the preparation of scaffolds by mixing acellular nucleus pulposus matrix and acellular cartilage matrix

Method used

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  • Nucleus pulposus-cartilage extracellular matrix scaffold and preparing method thereof
  • Nucleus pulposus-cartilage extracellular matrix scaffold and preparing method thereof
  • Nucleus pulposus-cartilage extracellular matrix scaffold and preparing method thereof

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Embodiment Construction

[0054] The present invention will be further described in conjunction with the accompanying drawings and specific embodiments.

[0055] 1. Preparation of decellularized nucleus pulposus matrix

[0056] The fresh sheep intervertebral disc nucleus pulposus tissue was rinsed with normal saline, soaked in 50mmol / L Tris-HCl buffer (buffer pH 7.5, containing 0.035mmol / L protease inhibitor - phenylmethylsulfonyl fluoride PMSF ), shaking on a shaking table (Beijing Liuyi Instrument Factory, WD-9405B type horizontal shaking table) at 4°C for 8 hours; crushed by a pulverizer (Shandong Joyoung Co., Ltd., pulverizing and grinding machine), and the diameter was obtained by density gradient centrifugation 100nm-5μm nucleus pulposus matrix fibrils; put into sterile Tris-HCl buffer solution (pH7.5) containing 0.6% TritonX-100 and 1.0% sodium deoxycholate, shake continuously at 4°C for 12h, Remove cell components; after thoroughly washed with sterile ultrapure water, put them into sterile PBS...

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Abstract

The invention discloses a nucleus pulposus-cartilage extracellular matrix scaffold and a preparing method thereof and belongs to the biological tissue engineering technology. The preparing method comprises the steps that nucleus pulposuses and bony articular surface cartilage are smashed, decellularized, subjected to gradient centrifugation to collect extracellular matrix microfilament suspension, and freeze-dried to obtain a freeze-dried decellularized nucleus pulposus matrix and a freeze-dried decellularized cartilage matrix, the two matrixes are taken in certain proportion and dissolved in ultrapure water to prepare mixed matrix suspension with certain concentration, and the mixed matrix suspension is freeze-dried and subjected to ultraviolet irradiation crosslinking and chemical reagent crosslinking in sequence so that the nucleus pulposus-cartilage extracellular matrix scaffold can be obtained. In this way, the defect that the nucleus pulposuses are hard to obtain due to the small number is overcome; the preparing technology is simple, and decellularizing can be achieved thoroughly; the matrix scaffold is degradable and high in biocompatibility; the matrix scaffold is of a three-dimensional structure, is high in mechanical property and plasticity, and can facilitate the distribution, growth and differentiation of seed cells; the freeze-dried extracellular matrixes are convenient to store and prepare and can be used for constructing a tissue engineered intervertebral disc to repair partial defects of the intervertebral disc, and broad clinical application prospects are realized.

Description

technical field [0001] The invention relates to biological tissue engineering technology, in particular to a nucleus pulposus-cartilage extracellular matrix scaffold and a preparation method thereof. Background technique [0002] With the aging of the population and changes in people's work and lifestyle, there are more and more degenerative disc diseases, especially neck and shoulder pain, low back and leg pain caused by disc herniation are very common, and severe cases often require surgical treatment. Resection is the most commonly used surgical method in spine surgery, but there has been no ideal repair and reconstruction method after disc resection. With the rapid development of tissue engineering technology, intervertebral disc tissue engineering has brought the possibility of permanent repair of intervertebral disc defects after surgery. [0003] In recent years, there have been many studies on scaffold materials used to construct tissue engineered intervertebral dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/36A61L27/56
Inventor 徐宝山杨强袁秋明马信龙张杨许海委
Owner TIANJIN HOSPITAL
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