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Implantable liposome embedded matrix composition, uses thereof, and polycaprolactone particles as scaffolds for tissue regeneration

a technology of embedded matrix composition and polycaprolactone, which is applied in the direction of phosphorous compound active ingredients, prosthesis, immunological disorders, etc., can solve the problems of changing the concentration of compound in the matrix and affecting the bioactivity of the compound

Inactive Publication Date: 2012-04-19
SCHUBERT HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0004]In exemplary embodiments, liposomes are introduced to a matrix (hydrogel) and are embedded in the matrix. The matrix can include one or more agents for recruiting cells (e.g., monocytes) to the matrix. Liposomes may encapsulate agents before being contacted with a matrix of the invention in some embodiments. In some embodiments, a matrix of the invention is formulated to incorporate agents and compounds that exert one or more effects upon cells which infiltrate the matrix. In other embodiments, a matrix of the invention is formulated to incorporate agents and compounds that attract cells

Problems solved by technology

However, a common problem of matrix based delivery systems is the diffusion of compounds embedded in the matrix into the surrounding tissue, changing the concentration of the compound in the matrix and affecting its bioactivity.

Method used

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  • Implantable liposome embedded matrix composition, uses thereof, and polycaprolactone particles as scaffolds for tissue regeneration

Examples

Experimental program
Comparison scheme
Effect test

example 1

Localized Differentiation of Endogenous Monocytes into an Angiogenic Phenotype

[0201]a. Preparation of Liposomes Containing IL-4 and IL-10 and Liposomes Containing Adenosine.

Reagents

Dioleoyl Phosphatidyl Choline

Saline (0.9% NaCl)

[0202]Human recombinant IL-4 and IL-10 are used together to activate the angiogenic phenotype in monocytes.

[0203]Adenosine can be used alone or in combination with IL-4 and IL-10 to activate the angiogenic phenotype in monocytes.

Method of Producing Liposomes Containing IL-4 and IL-10 for Targeting of Monocytes

Under Aseptic Conditions:

[0204]1. Dry 0.5 μmole of dioleoyl phosphatidyl choline under nitrogen in a disposable glass tube.

2. Evacuate in dessicator under vacuum for 30 minutes.

3. Add saline to required volume and scrape the sides of the glass tube to dislodge the lipid.

4. Add IL-4 and IL-10 1 μg / μl of lipid used, and, optionally adenosine 200 μM.

5. Vortex for 30 seconds. Sonicate twice in a bath sonicator at 7° C. for 1 min. This makes multilamellar ves...

example 2

Presentation of Cancer Cell Antigens to Monocytes / Dendritic Cells in the Vicinity of a Tumor

[0208]a. Preparation of Patient Specific Tumor Antigens

[0209]Tumor cells are taken by biopsy from a patient. The cells are lysed mechanically or by using a surface active material such as triton-X100. If surface antigens from the patient's cancer cells are required, cells can be incubated in a hypotonic buffer such as distilled water to remove cell content, and protein can then be extracted from the membranes. Protein extract from the cells is filtered to remove surface active materials and resuspended in saline solution. The protein extract from the cells is used for loading into liposomes. Proteins are loaded inside the liposomes to be phagocytosed by monocytes infiltrating the implanted matrix.

b. Preparation of the Liposomes Containing Tumor Antigens

Reagents

Dioleoyl Phosphatidyl Choline

Saline (0.9% NaCl)

Method

Under Aseptic Conditions:

[0210]1. Dry 0.5 μmole of dioleoyl phosphatidyl choline ...

example 3

Localized Reprogramming of Endogenous Monocytes or Stem Cells into Functional Insulin-Producing Beta Cells in the Pancreas

[0214]a. Preparation of Liposomes Containing Expression Vectors that Encode Transcription Factors

Reagents

Dioleoyl Phosphatidyl Choline

Saline (0.9% NaCl)

Method

Under Aseptic Conditions:

[0215]1. Dry 0.5 μmole of dioleoyl phosphatidyl choline under nitrogen in a disposable glass tube.

2. Evacuate in dessicator under vacuum for 30 minutes.

3. Add saline to required volume and scrape the sides of the glass tube to dislodge the lipid.

4. Add expression vectors that encode Ngn3, Pdx1 and Mafa, 0.1 μg / μl of lipid used.

5. Vortex for 30 seconds. Sonicate twice in a bath sonicator at 7° C. for 30 sec. This makes multilamellar vesicles that become small unilamellar vesicles (SUV) with prolonged sonication time. To make large unilamellar vesicles, an extruder can be used.

Another Method to Produce Liposomes Containing Transcription Factors for Targeting Monocytes:

[0216]Expression ...

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Abstract

In various embodiments, the present invention describes materials and methods for the local reprogramming of cells in a location where the treatment is applied. The invention can be used to replace lost cells or to restore function to tissue damaged due to disease, injury or genetic defect. In various embodiments, the treatment includes a semisolid hydrogel embedded with liposomes. The liposomes can contain an effector molecule or molecules. When phagocytic cells such as monocytes infiltrate the hydrogel, they encounter the liposomes and incorporate the liposomes carrying the effector molecules into the cells. In some embodiments, the effector molecules can be genetic material encoding the expression of specific proteins such as transcription factors, the expression of which can initiate the reprogramming of the cells. In other embodiments, the effector molecules can induce angiogenesis. In other embodiments, the effector molecules are tumor antigens. The matrix can contain other effector molecules designed to attract specific cells to the matrix. The cells can be released from the matrix as the matrix degrades or by active migration from the matrix. The cells can also remain in the matrix and secret molecules such as proteins and hormones that will diffuse through the matrix material to the surrounding tissue.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of priority from U.S. Ser. No. 61 / 200,208, filed Nov. 24, 2008; U.S. Ser. No. 61 / 200,213, filed Nov. 24, 2008; U.S. Ser. No. 61 / 200,207, filed Nov. 24, 2008; and 61 / 200,214, filed Nov. 24, 2008. The contents of the foregoing applications are incorporated herein by reference in their entirety.BACKGROUND OF THE INVENTION[0002]Delivery of bioactive molecules or chemical compounds to specific sites in vivo can be effected by embedding the molecules or compounds in a matrix. However, a common problem of matrix based delivery systems is the diffusion of compounds embedded in the matrix into the surrounding tissue, changing the concentration of the compound in the matrix and affecting its bioactivity.SUMMARY OF THE INVENTION[0003]The present invention is based on the development of compositions and methods for delivery of compounds encapsulated in liposomes. Embodiments of the invention feature liquid, semi-solid and solid matri...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K31/7052A61K31/7088A61K31/713A61K38/19A61K38/20A61K35/14A61K39/00A61P37/04A61K31/19A61K38/44A61K38/45A61K31/663A61P35/00A61P29/00A61P5/48A61K38/02
CPCA61K48/0016A61L2400/06A61L27/3804A61L27/48A61L27/54A61L2300/252A61L2300/258A61L2300/426A61L2300/45A61L2300/626A61L27/18A61L27/44A61L27/52A61L2300/232A61L27/24A61L27/222A61L2300/438C08L67/04A61P5/48A61P29/00A61P35/00A61P37/04
Inventor SCHUBERT, SHAI YEHOSHUA
Owner SCHUBERT HLDG
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