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Composition for Treatment of Damaged Part

a technology for treating damaged parts and parts, applied in the direction of skeletal/connective tissue cells, peptide/protein ingredients, peptide sources, etc., can solve the problems of morality and safety serious problems, the number and proliferation and differentiation potential of bone marrow stem cells (bmscs) decline with increasing, and the inability to perform invasive and painful procedures

Inactive Publication Date: 2013-08-01
UNIVERSITY OF TOKUSHIMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention provides a new way to treat damaged parts of the body using a type of dental stem cell. A method for making the treatment and using it to treat damaged parts is also provided. This treatment offers a new and effective way to treat damaged parts of the body.

Problems solved by technology

However, bone marrow aspiration is an invasive and painful procedure for the donor.
In addition, the number, and proliferation and differentiation potential of bone marrow stem cells (BMSCs) decline with increasing age.
Transplantation therapy of an intractable neurological disorder using neural stem cells from human embryos or ES cells (for example, Japanese Patent Application Laid-open (JP-A) No. 2002-281962) is recognized as a realistic research target, but has a serious problem in terms of morality and safety.
However, the result of the use of growth factors in single administration or multiple administrations, or the result of the use of multiple growth factors in combination with a view to obtaining synergistic effects, has not been clinically confirmed.

Method used

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  • Composition for Treatment of Damaged Part
  • Composition for Treatment of Damaged Part
  • Composition for Treatment of Damaged Part

Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials and Methods

[0233](1) Subjects and Cell Cultures

[0234]Human dental pulp tissues were obtained from clinically healthy extracted deciduous teeth and permanent teeth from eight patients. These experimental protocols were approved by the ethics committee of Nagoya University. SHED and DPSCs were isolated and cultured as described in Proc Natl Acad Sci USA 2000; 97: 13625-30 or Proc Natl Acad Sci USA 2003; 100: 5807-12.

[0235]Briefly, the pulp was gently removed and digested in a solution of 3 mg / mL collagenase type I and 4 mg / mL dispase at 37° C. for 1 hour. After filtration using 70-mm cell strainers (Falcon; BD Labware, Franklin Lakes, N.J.), cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; GIBCO, Rockville, Md.) containing 20% mesenchymal cell growth supplement (Lonza Inc., Walkersville, Md.) and antibiotics (100 U / mL penicillin, 100 mg / mL streptomycin and 0.25 mg / mL amphotericin B; GIBCO) at 37° C. with 5% CO2. After primary culture, the cells were subcultured...

example 2

(1) Preparation of Growth Factor Mixture (Powder)

[0270]Immortalized human mesenchymal stem cells (MSCs: Ronza Co., Ltd, USA) were used to prepare a growth factor (GF) admixture. The cells were cultured for 2 to 8 passages using 10% FSC-containing DMEM. At a stage of 80% confluence of cells in the culture dish, the supernatant (culture medium: CM) was sampled. The sampled CM was then added to ethanol (CM: Ethanol=1:9), and incubated at −20° C. for 60 min. The CM was concentrated by spinning (4° C., 1500 rpm, 15 min). The remaining CM was washed with 90% ethanol at −20° C., and then spun again.

[0271]The concentrated CM was freeze-dried, thereby obtaining a growth factor powder.

(2) Growth Factors in the Powder

[0272]Each growth factor in the powder was analyzed by the Western-blotting method. The detected growth factors are as follows: PDGF, VEGF, IGF, KGF, HGF and TGF.

example 3

Experimental Study of Bone Regeneration by Growth Factor

[0273](1) Methods

[0274]Four cylinder-shaped bone defects, each having a diameter of 10 mm and a depth of 10 mm, were formed in the dog mandible.

[0275]A titanium implant (3.75 mm in diameter) was inserted into the center of each defect.

[0276]The spaces around the implant were filled with graft materials, such as (1) PRP, (2) 100% GF, (3) MSCs (1×1,000,000) and (4) empty defect (control) (FIG. 10).

[0277](2) Results

[0278]Eight weeks later, the dog was euthanized and the mandible with the implants was dissected. A histological specimen was made and BIC (Bone-Implant Contact) was calculated using a light microscope and an image analyzing system (FIG. 11).

BIC=total length of bone contact to implant surface / total length of implant surface×100(%)

[0279]There was no significant difference in BIC between MSC (65.0) and GF (58.6), and the respective BIC values in MSC and GF were much higher than those of the control (26.4) and PRP (44.2) (...

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Abstract

The present invention provides a damaged part treatment composition for repairing a damaged part of a target tissue that includes a stem cell-conditioned medium obtained by culturing stem cells; a damaged part treatment method for repairing or restoring a damaged part of a target tissue that includes administering the damaged part treatment composition to a patient having the target tissue for the damaged part treatment composition in an amount therapeutically effective for repairing the damaged part of the target tissue; a method of treating cerebral infarction that includes administering the damaged part treatment composition to a cerebral infarct patient in an amount effective for repairing a damaged part of the brain; and a method of treating a CNS disease that includes administering, as a CNS disease treatment composition, the damaged part treatment composition to a CNS disease patient in a therapeutically effective amount.

Description

TECHNICAL FIELD[0001]The present invention relates to a composition for treatment of a damaged part, and a treatment method using the same.[0002]Regenerative medicine utilizing stem cells attracts attention as a versatile alternative technique for diseases that are hard to treat by conventional medicine.[0003]Regenerative medicine using stem cells is a promising tool in a new clinical platform for a whole spectrum of intractable diseases. Various stem cells have been reported, including embryonic stem cells (ES cells), induced pluripotent stem cells (iPS cells) and somatic stem cells. Among somatic stem cells, mesenchymal stem cells (MSCs) isolated from various tissues including bone marrow, adipose tissue, skin, umbilical cord and placenta have been used in particular in clinical applications in skin regeneration. However, bone marrow aspiration is an invasive and painful procedure for the donor. In addition, the number, and proliferation and differentiation potential of bone marro...

Claims

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Application Information

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IPC IPC(8): A61K35/12A61K35/28A61K35/30A61K35/32A61K35/545
CPCA61K35/545A61K38/1833C12N5/0664A61K38/1866A61K38/18A61K35/28A61K35/30A61K35/32A61K38/30A61K38/1841A61K38/1858A61K35/12A61K2300/00A61P1/02A61P17/00A61P17/02A61P19/08A61P25/00A61P25/14A61P25/16A61P25/28A61P27/02A61P9/10
Inventor UEDA, MINORUYAMADA, YOICHIEBISAWA, KATSUMIYAMAMOTO, AKIHITOSAKAI, KIYOSHIMATSUBARA, KOHKIHATTORI, HISASHISUGIYAMA, MASAHIKOINOUE, TAKANORI
Owner UNIVERSITY OF TOKUSHIMA
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