Vortex-induced silk fibroin gelation for encapsulation and delivery

a technology of silk fibroin and encapsulation, which is applied in the direction of antibody medical ingredients, peptide/protein ingredients, drug compositions, etc., can solve the problems of limiting cell viability and altering cell or bioactive molecule function

Inactive Publication Date: 2012-03-22
TRUSTEES OF TUFTS COLLEGE TUFTS UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0006]Some embodiments of the invention provide for methods of controlling the gelation time of silk fibroin initiated by vortexing a silk fibroin solution for a sufficient period of time to initiate gelation. For examp

Problems solved by technology

Traditional gelation methods using aqueous native silk protein solutions, under physiologically relevant conditions, often range from days to weeks for gelation: too slow for the incorporation of cells and labile

Method used

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  • Vortex-induced silk fibroin gelation for encapsulation and delivery
  • Vortex-induced silk fibroin gelation for encapsulation and delivery
  • Vortex-induced silk fibroin gelation for encapsulation and delivery

Examples

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example 1

Preparation of Aqueous Silk Fibroin Solutions

[0111]Silk fibroin aqueous solutions were prepared as previously described in the literature. See Sofia et al., 54 J. Biomed. Mater. Res. 139-48 (2001). Briefly, Bombyx mori cocoons were boiled for 40 min in an aqueous solution of 0.02 M NaCo3 and then rinsed thoroughly with deionized water. After overnight drying, the extracted silk fibroin was dissolved in an aqueous solution of 9.3 M LiBr at 60° C. overnight. The resulting solution was dialyzed against deionized water using Slide-A-Lyzer dialysis cassettes (MWCO 3,500, Pierce, Thermo Scientific, Waltham, Mass.) for two days to remove the residual salt. The final concentration of the silk fibroin was approximately 5.3 wt %. Lower concentration silk solutions were prepared by diluting the 5.3 wt % stock solution with deionized water. Additionally, the silk fibroin solution may be concentrated, for example, to about 30% (w / v). Briefly, the silk fibroin solution with a lower concentration ...

example 2

Vortex-Induced Hydrogelation

[0113]A 1 ml aliquot of silk solution was equilibrated at 25° C. in a vial kept in a water bath for 10 min and mixed for predetermined times at 3,200 rpm using a vortexer (Fisher Scientific, Pittsburg, Pa.) to induce silk self-assembly and hydrogelation. Such prepared silk hydrogelation was characterized by CD and rheology experiment. Increasing the vortex time increased the solution turbidity and eventually bulk phase separation of a white and solid-like material was observed, especially at lower protein concentrations. Both CD and rheology data were collected from turbid solutions after removal of the solid phase.

example 3

Circular Dichroism Spectroscopy

[0114]Circular dichroism (CD) spectra were collected using an Aviv Model 410 CD spectrometer (Aviv Biomedical, Inc., Lakewood, N.J.). After vortexing, aqueous silk solutions were immediately loaded in a 0.1 mm quartz cell within a temperature controlled cell holder. CD wavelength scans between 210 nm and 260 nm or time sweeps at 216 nm were collected at 25° C. Due to the high silk concentrations, the high dynode voltages below 210 nm lead to erroneous data. The CD spectrum of water collected immediately before each measurement was used for background correction.

[0115]The mean residual ellipticity was calculated from

[θ]=θ·M10·c·l·n(deg·cm2dmol)

where θ is the measured ellipticity (deg), M is the mean molecular mass (g / mol), c is the protein concentration (g / cm3), 1 is the path length (cm), and n is the number of residues. M and n for B. mori heavy chain were taken as 391,563 g / mol and 5263, respectively (Zhou et al, 28 Nucleic Acids Res. 2413-19 (2000))....

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Abstract

The present invention provided for a novel process of forming silk fibroin gels, and controlling the rate of β-sheet formation and resulting hydrogelation kinetics, by vortex treatment of silk fibroin solution. In addition, the vortex treatment of the present invention provides a silk fibroin gel that may be reversibly shear-thinned, enabling the use of these approach for precise control of silk self-assembly, both spatially and temporally. Active agents, including biological materials, viable cells or therapeutic agents, can be encapsulated in the hydrogels formed from the processes, and be used as delivery vehicles. Hence, the present invention provide for methods for silk fibroin gelation that are useful for biotechnological applications such as encapsulation and delivery of active agents, cells, and bioactive molecules.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61 / 182,794 filed Jun. 1, 2009, and U.S. Provisional Application No. 61 / 219,952 filed Jun. 24, 2009, the contents of which are incorporated herein by reference in their entirety.GOVERNMENT SUPPORT[0002]This invention was made with government support under contract No. P41 EB002520 awarded by the National Institutes of Health, and No. FA9550-07-1-0079 awarded by the Air Force Office of Scientific Research. The U.S. government has certain rights in the invention.FIELD OF THE INVENTION[0003]This invention provides for methods of forming silk fibroin gels comprising vortexing, and methods of delivering active agents encapsulated in vortex-induced silk hydrogels.BACKGROUND OF THE INVENTION[0004]Biocompatible and biodegradable polymer hydrogels are useful carriers for delivering active agents and cells for biomedical applications, such as ...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K39/395A61K38/18A61K39/00A61K38/43A61K35/76A61K35/12A61K35/34A61K35/23A61K35/407A61K35/28A61K35/54A61K35/48A61P31/00A61K35/00
CPCA61K9/0024A61K38/00A61K47/42A61P31/00A61K9/06A61K9/5052A61K9/5089
Inventor KAPLAN, DAVID L.YUCEL, TUNA
Owner TRUSTEES OF TUFTS COLLEGE TUFTS UNIV
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