Materials and methods for augmenting and/or repairing intervertebral discs

A technology of intervertebral disc and human intervertebral disc, applied in the direction of biochemical equipment and methods, vertebrate cells, bone/connective tissue cells, etc.

Inactive Publication Date: 2006-05-31
WARSAW ORTHOPEDIC INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to date, such multipotent or pluripotent cells have never been used to expand or repair intervertebral discs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Obtaining Stem Cell Material from Somatic Tissue

[0067] Raw liposuction aspirate can be obtained from patients undergoing elective surgery. Before liposuction, the patient may be given epinephrine to minimize blood contamination of the aspirate. The aspirate is then subjected to coarse filtration to separate the associated adipose tissue mass from the associated liquid waste. The isolated tissue was rinsed with neutral phosphate buffered saline and then enzymatically digested with 0.075% w / v collagenase at 37°C for about 20 minutes with intermittent agitation.

[0068] After digestion the collagenase was inactivated and the slurry was centrifuged at about 260g for 10 minutes. This yielded multiple layers of supernatant and a cell pellet. The supernatant was then removed and saved for future use. The cell pellet was resuspended in a solution of lysed red blood cells and incubated at about 25°C for about 10 minutes without agitation. After the i...

Embodiment 2

[0072] Dedifferentiation of somatic cells into pluripotent stem cells in vitro

[0073]Adult keratinocytes were obtained from Clonetics (San Diego, CA) and grown in keratinocytes at 37°C, 5-10% CO2 according to the instructions in "Keratinocyte System Instructions" (BioWhittaker, catalog number AA-1000) grow in medium. When adult keratinocytes were approximately 40-80% confluent, 10-25 D M 5-aza-2'-deoxycytidine (Sigma, St Louis, MO) was added to the cultures. After 4 days of incubation with 5-aza-2'-deoxycytidine, 100-250 ng / ml trichophyllin (Sigma, St Louis, MO) was added to the culture. Cultures were grown for an additional day and samples were taken for telomerase activity.

[0074] Determination of telomeres in keratinocytes exposed to 10-25 D M 5-aza-2'-deoxycytidine for 5 days and 100-250 ng / ml trichostatin for 1 day, as well as cells not exposed to these agents Enzyme expression (using a commercially available TRAP assay kit). These cells ...

Embodiment 3

[0076] Growth of stem cells in vitro

[0077] Stem cells at 37 °C and 5% CO 2 cultured in DMEM supplemented with 10% fetal bovine serum. Under these conditions, cells can be passaged at least 5 times without differentiation and without losing their developmental phenotype.

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Abstract

A method of expanding and / or repairing an intervertebral disc by applying stem cell material in the disc. Stem cells can be undifferentiated cells, or they can be cells that have differentiated but subsequently dedifferentiated. Stem cells can be induced to express at least one characteristic of human intervertebral disc cells, such as fibroblasts, chondrocytes or dorsal cells, by exposing them to agents and / or environments that induce the desired differentiation. In some embodiments, the stem cell material can be provided along with a collagen-based material, which can be a collagen-rich lattice. Stem cell material can be provided as a stem cell isolate, which can be substantially free of non-stem cell material. Other therapeutic agents can be administered with the stem cell material.

Description

field of invention [0001] The present invention generally relates to materials and methods for expanding and / or repairing intervertebral discs, and more particularly to materials and methods for expanding and / or repairing intervertebral discs using stem cell materials. Background of the invention [0002] Healthy intervertebral discs facilitate movement between paired vertebrae while absorbing and distributing shock. The intervertebral disc consists of two parts: a soft central nucleus (nucleus pulposus) that bears most of the load and a tough outer ring (annulus fibrosus) that contains and stabilizes the core material. [0003] As the natural aging process progresses, the discs can dehydrate and degenerate, negatively affecting their ability to adequately cushion and support the vertebral body. This natural shrinkage is often referred to as a "black disc" in more advanced stages, as dehydration of the disc appears on magnetic resonance imaging [MRI] and the vertebrae can m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61F2/00A61F2/02A61F2/28A61F2/30A61F2/44C12N5/0775
CPCA61F2/442A61F2002/2817A61F2002/30062A61F2002/3008A61F2002/30677A61F2002/444A61F2002/4445A61F2002/445A61F2210/0004A61F2250/0098A61F2310/00365A61L27/3834A61L27/3856A61L27/3895A61L2430/38C12N5/0667C12N2500/25C12N2501/105C12N2501/115C12N2501/135C12N2501/15C12N2501/155C12N2501/39
Inventor H·H·特里优
Owner WARSAW ORTHOPEDIC INC
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