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Electrospun cell matrices

a technology of electrospun cells and matrices, which is applied in the field of cellular attachment and growth matrices, can solve the problems of eliciting adverse immune responses in patients and limited supply, and achieves the effects of enhancing the functionality of the scaffold, high molecular weight, and fiber siz

Inactive Publication Date: 2006-09-14
WAKE FOREST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0006] Electrospinning matrices also allows control over the composition, geometry, and mechanics of cellular scaffolds. Electromagnetic fields created by high voltage sources during electrospinning cause biomaterials in a solvent to elongate and splay into ever-smaller fibers. As the fibers become smaller, evaporation of the solvent occurs more quickly and fiber sizes of nanometer diameter can be achieved. The fibers are drawn to a grounded surface or will adhere to surfaces placed between the solution source and ground. This process also offers the capability to incorporate nanomaterials to enhance the functionality of the scaffold.
[0007] In one aspect of the invention, the electrospun matrices comprise at least one natural biological material component and at least one synthetic polymer material. In one embodiment, the natural biological material component further comprises collagen (which can be derived from biological tissue sources or synthesized) and the synthetic polymer component can a high molecular weight polymer, e.g., having a molecular weight of at least 1000, preferably between 2,000 and 20,000. The natural component makes the matrices highly biocompatible and / or less immunogenic while the molecular weight polymer component can impart additional mechanical strength to the matrix and / or improve ease of manufacture by increasing viscosity and spinning characteristics of the solution during electrospining. The polymeric component can be, for example, a poly(lactic acid) polymer or a poly(glycolic acid) polymer or a co-polymer thereof, such as poly(D,L-lactide-co-glycolide), with a molecular weight from about 1000 to about 20,000. The materials being used in the electrospun matrice are capable of being charged or capable of being transported in a charged solution.
[0008] The electrospun matrices can further comprise a second natural component chosen to impart elasticity. For example, the natural biological material component can comprise elastin (which again can be derived from biological tissue sources or synthesized). Following electrospining, the matrix can also be crosslinked for increased stability and strength, using various known crosslinking methods.

Problems solved by technology

However, these synthetic matrices often elicit an adverse immune response in a patient.
While decellularized matrices are a promising as scaffolds, they are only available in a limited supply.

Method used

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Examples

Experimental program
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Effect test

example 1

Methods and Materials

(i) Scaffold Preparation

[0148] Electrospun nanofiber scaffolds have been developed using a solution of collagen type I, elastin, and poly(D,L-lactide-co-glycolide) (PLGA, mol. ratio 50:50, Mw 110,000) (Boeringer-Ingelheim, Germany). Collagen type I from calf skin (Elastin Products Company, Owensville, Mo.), elastin from ligamentum nuchae (bovine neck ligament), (Elastin Products Company, Owensville, Mo.), and PLGA are mixed at a relative concentration by weight of 45% collagen, 40% PLGA, and 15% elastin. The solutes are dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (99+%) (Sigma Chemical Company, St. Louis, Mo.) at a total solution concentration of 15 w / v % (150 mg / mL). High molecular weight PLGA, previously used for electrospinning tissue scaffolds is added to the solution to increase mechanical strength of the scaffold and increase viscosity and spinning characteristics of the solution.

[0149] Physically, the electrospinning method requires a high voltage ...

example 2

Preparation of Decellularized Organs

[0167] The following method describes a process for removing the entire cellular content of an organ or tissue without destroying the complex three-dimensional infra-structure of the organ or tissue. An organ, e.g. a liver, was surgically removed from a C7 black mouse using standard techniques for tissue removal. The liver was placed in a flask containing a suitable volume of distilled water to cover the isolated liver. A magnetic stir plate and magnetic stirrer were used to rotate the isolated liver in the distilled water at a suitable speed for 24-48 hours at 4° C. This process removes the cellular debris and cell membrane surrounding the isolated liver.

[0168] After this first removal step, the distilled water was replaced with a 0.05% ammonium hydroxide solution containing 0.5% Triton X-100. The liver was rotated in this solution for 72 hours at 4° C. using a magnetic stir plate and magnetic stirrer. This alkaline solution solubilized the nuc...

example 3

Electrospun Matrices

[0171] An electrospun matrix was formed using the methods outlined in Example 1. A solution of collagen type I, elastin, and PLGA, were used. The collagen type I, elastin, and PLGA were mixed at a relative concentration by weight of 45% collagen, 40% PLGA, and 15% elastin.

[0172] The resulting fibrous scaffold had a length of 12 cm with a thickness of 1 mm. A 2 cm representative sample is depicted in FIG. 3. This demonstrates the feasibility of spinning Type I Collagen and elastin into fibers from nanometer to micrometer diameter using concentrations from 3% to 8% by weight in solution. These results also show that by adding PLGA (Mw 110,000) to the mixture, solutions with higher viscosity and improved spinning characteristics could attained. By increasing the solution concentration to 15%, thicker, stronger scaffolds were able to be built while maintaining the collagen and elastin components.

[0173] Collagen type I stained positively on the decellularized scaff...

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Abstract

The invention is directed to compositions and methods for preparing electrospun matrices comprising at least one natural biological material component and at least one synthetic polymer material. The natural component makes the matrices highly biocompatible while the molecular weight polymer component can impart additional strength mechanical strength to the scaffold and / or improve ease of manufacture by increasing viscosity and spinning characteristics of the solution during electrospining.

Description

RELATED APPLICATION [0001] The present application claims priority to a provisional application entitled “Electrospun Cell Matrices” filed on Mar. 11, 2005 and having serial no. ______ (Atty Dkt No. 105447-3).BACKGROUND OF THE INVENTION [0002] The technical field of this invention relates to matrices for cellular attachment and growth. The invention also relates to methods of making and using these matrices for tissue engineering and the construction of artificial structures. [0003] Synthetic matrices have been used for artificial tissue construction. However, these synthetic matrices often elicit an adverse immune response in a patient. To circumvent this problem, decellularized matrices have been used. These decellularized matrices are advantageous for several reasons. They are naturally derived, and therefore less likely to induce an adverse immune response. They also have a similar composition, ultrastructure and biomechanics to the native tissue. While decellularized matrices a...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61F2/02
CPCD01D5/0007D01F4/00C12N2533/54C12N5/0691C12N2533/40D01F6/92
Inventor ATALA, ANTHONYYOO, JAMESLIM, GRACECZERW, RICHARDSOKER, SHAYSTITZEL, JOEL
Owner WAKE FOREST UNIV
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