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Bone Augmentation Utilizing Muscle-Derived Progenitor Compositions in Biocompatible Matrix, and Treatments Thereof

a technology of biocompatible matrix and progenitor cells, applied in the field of muscle-derived progenitor cells, can solve the problems of low survival rate of cells following transplantation, association of primary myoblast-derived treatments, and polymer solution

Inactive Publication Date: 2015-02-12
UNIVERSITY OF PITTSBURGH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In spite of the above, in most cases, primary myoblast-derived treatments have been associated with low survival rates of the cells following transplantation due to migration and / or phagocytosis.
However, the polymer solution presents the same problems as the biopolymers discussed above.
Furthermore, the Atala patent is limited to uses of myoblasts in only muscle tissue, but no other tissue.
It is notable that prior attempts to use myoblasts for non-muscle tissue augmentation were unsuccessful (U.S. Pat. No. 5,667,778 to Atala).
While SIS is primarily used for the repair of soft tissues, its potential as a bone graft material is still under debate.
J Biomed Mater Res, 2004 suggests that SIS is not capable of inducing or conducting new bone formation across a critical size segmental bone defect.
Moreover, current methods of producing cell matrices for in vivo tissue and organ repair are very costly and time consuming.
Such cell matrices are costly due to the specialized factories and / or procedures needed to produce these products.
Also, since cell-matrix products involve living biological cells / tissue, a tremendous loss of product occurs from shipping, the delays associated therewith, and the like.
Additionally, given the nature of the products, obtaining regulatory approval for new products that are based on living cells and a new matrix poses difficulties.
Those in the art have recognized that a major problem remaining to be solved is the delay in producing the cell-matrix product after initial preparation.
Specifically, it has been stated that there is a problem of a three week delay necessary to produce a sufficient amount of autologous keratinocytes and fibroblasts for the production of reconstructed skin.

Method used

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  • Bone Augmentation Utilizing Muscle-Derived Progenitor Compositions in Biocompatible Matrix, and Treatments Thereof
  • Bone Augmentation Utilizing Muscle-Derived Progenitor Compositions in Biocompatible Matrix, and Treatments Thereof
  • Bone Augmentation Utilizing Muscle-Derived Progenitor Compositions in Biocompatible Matrix, and Treatments Thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

MDC Enrichment, Isolation and Analysis According to the Pre-Plating Method

[0075]MDCs were prepared as described (U.S. Pat. No. 6,866,842 of Chancellor et al.). Muscle explants were obtained from the hind limbs of a number of sources, namely from 3-week-old mdx (dystrophic) mice (C57BL / 10ScSn mdx / mdx, Jackson Laboratories), 4-6 week-old norrnal female SD (Sprague Dawley) rats, or SCID (severe combined immunodeficiency) mice. The muscle tissue from each of the animal sources was dissected to remove any bones and minced into a slurry. The slurry was then digested by 1 hour serial incubations with 0.2% type XI collagenase, dispase (grade II, 240 unit), and 0.1% trypsin at 37° C. The resulting cell suspension was passed through 18, 20, and 22 gauge needles and centrifuged at 3000 rpm for 5 minutes. Subsequently, cells were suspended in growth medium (DMEM supplemented with 10% fetal bovine serum, 10% horse serum, 0.5% chick embryo extract, and 2% penicillin / streptomycin). Cells were then...

example 2

MDC Enrichment, Isolation and Analysis According to the Single Plate Method

[0079]Populations of rapidly- and slowly-adhering MDCs were isolated from skeletal muscle of a mammalian subject. The subject may be a human, rat, dog or other mammal. Biopsy size ranged from 42 to 247 mg.

[0080]Skeletal muscle biopsy tissue is immediately placed in cold hypothermic medium (HYPOTHERMOSOL® (BioLife) supplemented with gentamicin sulfate (100 ng / ml, Roche)) and stored at 4° C. After 3 to 7 days, biopsy tissue is removed from storage and production is initiated. Any connective or non-muscle tissue is dissected from the biopsy sample. The remaining muscle tissue that is used for isolation is weighed. The tissue is minced in Hank's Balanced Salt Solution (HBSS), transferred to a conical tube, and centrifuged (2,500×g, 5 minutes). The pellet is then resuspended in a Digestion Enzyme solution (Liberase Blendzyme 4 (0.4-1.0 U / mL, Roche)). 2 mL of Digestion Enzyme solution is used per 100 mg of biopsy t...

example 3

Small Intestine Submucosa Alleviates the Repair of a Critical Size Calvarial Defect in Mice

[0085]The purpose of this study was to investigate the bone regenerative potential of single-layer SIS scaffold transplanted into critical size calvarial defect in mice. We also preconditioned SIS grafts by seeding them with human muscle-derived cells (hMDCs), prepared as detailed in Example 2, above, in order to test osteogenic potential of this construct in response to natural fracture environment.

Materials and Methods

[0086]In this study a total of 24 SCID mice were used. All animal experiments were approved by institutional ARCC. Surgical procedure was performed under general anesthesia. Critical size calvarial bone defect was created using a 5-mm-diameter trephine burr. Human muscle-derived cells (hMDCs) isolated from a 35 year old male patient were provided. Animals were divided into 3 groups according to the treatment they received. A control group consisted of untreated mice with a calv...

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Abstract

The present invention provides muscle-derived progenitor cells that show long-term survival following transplantation into body tissues and which can augment non-soft tissue following introduction (e.g. via injection, transplantation, or implantation) into a site of non-soft tissue (e.g. bone) when combined with a biocompatible matrix, preferably SIS. The invention further provides methods of using compositions comprising muscle-derived progenitor cells with a biocompatible matrix for the augmentation and bulking of mammalian, including human, bone tissues in the treatment of various functional conditions, including osteoporosis, Paget's Disease, osteogenesis imperfecta, bone fracture, osteomalacia, decrease in bone trabecular strength, decrease in bone cortical strength and decrease in bone density with old age.

Description

RELATED APPLICATIONS[0001]This application claims priority to and is a continuation of U.S. patent application Ser. No. 12 / 543,311, filed on Aug. 18, 2009, which claims priority from U.S. Provisional Patent Applications 61 / 089,798, filed on Aug. 18, 2008 and 61 / 166,775, filed on Apr. 6, 2009, incorporated by reference, herein, in their entireties.GOVERNMENT INTERESTS[0002]This invention was made with Government support under Grant No. R01-DE13420-01 awarded by the National Institutes of Health. The Government has certain rights in this invention.FIELD OF THE INVENTION[0003]The present invention relates to muscle-derived progenitor cells (MDCs) and compositions of MDCs with biologically compatible matrix and their use with the augmentation of body tissues, particularly bone. In particular, the present invention relates to muscle-derived progenitor cells that show long-term survival following introduction into bone used in combination with small intestine sub-mucosa for the augmentati...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61L27/38C12N11/02A61L27/36A61L27/54A61L27/58A61K35/12A61K35/38
CPCA61L27/3895A61L27/54A61L27/58A61L27/3847A61L2300/64A61L27/3629C12N11/02A61L2430/02A61L2430/40A61L27/365A61K35/12A61K35/38C12N5/0659C12N2533/92A61L27/3826A61P19/00A61P19/08A61P19/10C12N5/0652C12N5/0658
Inventor USAS, ARVYDASPAYNE, KARINPAYNE, THOMASJANKOWSKI, RONALDHUARD, JOHNNY
Owner UNIVERSITY OF PITTSBURGH
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