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Treatment of collagen defects using protein solutions

a technology of collagen and protein solution, which is applied in the direction of peptide/protein ingredients, peptide sources, drug compositions, etc., can solve the problems of genetic defects or nutritional deficiencies, wear, trauma or disease, and may be debilitating and painful

Inactive Publication Date: 2014-09-18
BIOMET BIOLOGICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present technology provides methods and compositions for stimulating the production of collagen, specifically by increasing the production of chondrocytes. The methods involve administering a blood-derived composition containing specific proteins to the site of a collagen defect. The composition may also contain cytokine-producing cells, platelets, or a combination of both. The technology also provides methods for making the compositions by obtaining a cytokine cell suspension from the subject and fractionating it to produce an autologous protein solution. The autologous protein solution is then administered to the collagen defect site. The technical effects of the technology include improved collagen production and reduced inflammation at the site of collagen defects.

Problems solved by technology

Diseases may result from genetic defects or nutritional deficiencies that affect the production of collagen by chondrocytes.
Such diseases may be debilitating and painful.
Cartilage defects can appear as a hole or a tear in a cartilage surface and can result from wear, trauma or disease.
Since cartilage has minimal ability to repair itself, even a small cartilage defect, if left untreated, can hinder the ability of a human or other mammal to move free from pain and can cause deterioration of a joint surface.
However, many such treatments may present side effects, and may have limited long term utility as underlying conditions become worse.

Method used

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  • Treatment of collagen defects using protein solutions
  • Treatment of collagen defects using protein solutions
  • Treatment of collagen defects using protein solutions

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparing and Characterizing a Protein Solution

[0163]A Protein Solution rich in interleukin-1 receptor antagonist is prepared from seven consented human providers. Blood (55 mL) is drawn into a 60 cc syringe with 5 mL of anticoagulant citrate dextrose solution A (ACD-A, Citra Labs, Braintree, Mass.). Platelet-rich plasma (PRP) is created using the GPS® III platelet concentration system (800-1 003A, Biomet Biologics, Warsaw, Ind.) according to the instructions for use. The solution is generated by adding 6 mL of PRP to a modified Plasmax device containing 1 gram of polyacrylamide beads (Biomet Biologics, Warsaw, Ind.). The IL-Ira solution is removed from the Plasmax devices and frozen at minus 50° C. for the assay. Cytokine content is assayed on a 16-plex ELISA (Searchlight Protein Array, Aushon Biosystems, Billerica, Mass.). The analytes included IL-4, IL-10, IL-11, IL-13, IL-Ira, IFN-γ, sTNF-RI, sTNF-RII, IL-1α, IL-1β, TNF-α, IL-17, IL-18, bFGF, TBF-β1, and TBF-β2.

[0164]The solutio...

example 2

Generation of IL-1ra from Platelet-Rich Plasma

[0165]An IL-1ra-rich solution is created as follows. Whole blood (70 mL) anticoagulated (10%) with ACD-A (Braintree, Mass., USA) is drawn from 5 healthy volunteers. A portion (10 mL) is reserved for a whole blood measurement. Platelet-rich plasma (PRP) (6 mL) is produced using the GPS® II System (Biomet Biologics, LLC, Warsaw, Ind., USA). Complete blood counts are collected for the whole blood and PRP samples following a validated procedure, as described in Woodell-May J E, Ridderman D N, Swift M J, Higgins J. “Producing Accurate Platelet Counts for Platelet Rich Plasma: Validation of a Hematology Analyzer and Preparation Techniques for Counting” J. Craniofac. Surg. (2005) September. 16(5):749-56.

[0166]Following the PRP production, 5 mL of the PRP is added to a modified plasma concentration device (Plasmax™, Biomet Biologics LLC, Warsaw, Ind., USA) and incubated with polyacrylamide desiccating beads in the device for 24 hours at room tem...

example 3

Production of Protein Solution from PRP

[0170]Anticoagulated blood (120 cc) is collected from 5 human donors. Platelet-rich plasma (PRP) is prepared using GPS® III disposables (Biomet Biologics LLC, Warsaw, Ind., USA). PRP is loaded into modified plasma concentration devices (Plasmax™, Biomet Biologics LLC, Warsaw, Ind., USA) and processed. The output is divided into 4 groups: IL-1ra in concentrated plasma with and without thrombin activation (1000 U / mL in 1M CaCl2), or cell-free IL-1ra with and without thrombin activation. IL-1ra is measured using ELISA (R&D Systems) over time.

[0171]The PRP contacts polyacrylamide beads in the Plasmax™ device while electromagnetic field stimulation is provided using a capacitively coupled electromagnetic field.

[0172]Unclotted PRP produces an average of about 50 ng over 24 hrs. The cell-free samples produce about 34 ng without changing over 24 hrs. Once clotted, the elution of IL-1ra is slowed, with only about 30% being eluted after 10 hours. Release...

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Abstract

Methods of stimulating collagen production, including stimulation of chondrocyte production, at the site of a defect. Methods include administering to the site of a defect at least two proteins from the group IL-1ra, sTNF-RI, sTNF-RII, IGF-I, EGF, HGF, PDGF-AB, PDGF-BB, VEGF, TGF-β1, and sIL-1RII.

Description

INTRODUCTION[0001]The present technology relates to methods of stimulating chondrocytes at the site of a collagen defect in mammalian subjects. In particular, methods comprise use of solutions comprising cytokines, including such solutions derived from blood and other tissues.[0002]Chondrocytes are cells that produce and maintain the cartilaginous matrix that comprises cartilage. Collagen is the most abundant protein in mammals, and is present in a variety of tissues, including skin, connective tissue, cartilage, bone, blood vessels and intervertebral disks. There are a variety of collagen-related disorders, resulting from disease as well as trauma to tissues comprising collagen. Diseases may result from genetic defects or nutritional deficiencies that affect the production of collagen by chondrocytes. Some collagen disorders as associated with autoimmune diseases, such as rheumatoid arthritis, lupus, and systemic sclerosis, which destroy collagen after it is produced. Such diseases...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/20A61K45/06A61K35/28A61K38/17A61K35/14
CPCA61K38/2006A61K35/19A61K45/06A61K38/1793A61K35/28A61K38/19A61K38/20A61K35/14A61P19/02A61K38/1808A61K38/1833A61K38/1858
Inventor MATUSKA, ANDREAO'SHAUGHNESSEY, KRISTAWOODELL-MAY, JENNIFER E.
Owner BIOMET BIOLOGICS
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