Support Having Nanostructured Titanium Dioxide Film And Uses Thereof

• US 20110229579A1
• Filed: 05/27/2011
• Published: 09/22/2011
• Est. Priority Date: 07/21/2005
• Status: Abandoned Application

• Current Assignee: Tethis S.p.A.
• Original Assignee: Tethis S.p.A.
• Inventors: Pelicci, Pier Giuseppe, Milani, Paolo, Barborini, Emanuele, Bongiorno, Gero Antonio, Piseri, Paolo, Carbone, Roberta
• Application Number:
  US13/117,973
• Publication Number:
  US 20110229579A1
• Time in Patent Office: Days
• US Class Current: 424/490
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: G01N 33/551 the carrier being inorganic
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

The present invention relates to supports for bioassays and the use thereof in cell culturing and in cell-based methods and assays. More precisely, the invention provides solid materials coated with films of nanostructured titanium dioxide suitable for the immobilisation of viruses and for cell-adhesion. The nanostructured TiO₂ film-coated support of the invention is particularly useful for the preparation of microarrays for genetic and phenotypic analysis.

Claims

1. A solid support fabricated from a biocompatible substrate material which is at least partially coated with a nanostructured TiO₂ film having virions and/or cells immobilised on the surface thereof.

2. The solid support of claim 1 wherein the film of nanostructured TiO₂ consists of TiO₂ nanoparticles with a diameter below 20 nm embedded in an amorphous TiO₂ matrix with a density of below 75% of bulk TiO₂ density.

3. The solid support of claim 1 which comprises a slide, dish, flask, plate, coverslip, fiber, foam, particle, membrane, porous scaffold, mesh or implant.

4. The solid support of claim 1 wherein the biocompatible substrate material is glass, plastic, ceramic, metal or a biodegradable or undegradable biopolymeric material.

5. The solid support of claim 1 wherein the virions are retroviruses, adenoviruses, adeno-associated viruses (AAV) or any other viruses that can be utilized as vectors for genetic manipulation of cells.

6. The solid support of claim 1 wherein the virions are genetically modified.

7. The solid support according to claim 1 wherein the nanostructured TiO₂ film includes or is coated with streptavidin, avidin or neutravidin, and biotinylated virions.

8. The solid support according to claim 1 wherein the nanostructured TiO₂ film is in the form of a micro- or nano-pattern.

9. The solid support according to claim 8 wherein virions carrying different genetic inserts are spotted on the surface of the nanostructured TiO₂ film.

10. A method for cell infection with virus in vitro comprising the steps of:

(a) providing a solid support of a biocompatible substrate material which is at least partially coated with a nanostructured TiO₂film; and

(b) culturing cells on the nanostructured TiO₂ film-coated support in the presence of an infecting virion.

11. The method of claim 10 comprising the further steps of:

(c) contacting the nanostructured TiO₂ film-coated support with virions prior to culturing cells in the support;

(d) contacting the virion adhered on the surface of the nanostructured TiO₂film-coated support with a cell preparation; and

(e) culturing the cells for a time sufficient for the infection to occur.

14. The method of claim 10 wherein the virions are retroviruses, adenoviruses, adeno-associated viruses (AAV) and any other viruses that can be utilized as vectors for genetic manipulation of cells.

15. The method of claim 10 wherein the virions are genetically modified.

12. A method for cell infection with viruses in vitro comprising the steps of:

(a) providing a solid support of a biocompatible substrate material which is at least partially coated with a nanostructured TiO₂ film;

(b) immobilising streptavidin, avidin or neutravidin on the nanostructured TiO₂ film coating;

(c) contacting the nanostructured TiO₂ film-coated support with a biotinylated virion so as to form a complex of biotinylated virions with the immobilised streptavidin, avidin or neutravidin;

(d) contacting the complex with a cell preparation; and

(e) culturing the cells for a time sufficient for the infection to occur.

13. A method for cell infection with viruses in vitro comprising the steps of:

(a) providing a solid support of a biocompatible substrate material being at least partially coated with a nanostructured TiO₂ film;

(b) adding a cell preparation to the nanostructured TiO₂ film-coated support;

(c) culturing the cells for an appropriate period of time;

(d) adding a virion supernatant;

(e) culturing the cells for a time sufficient for the infection to occur.

16. A method for gene therapy ex vivo comprising the steps of:

(a) recovering cells to be genetically modified from a patient;

(b) establishing a primary cell culture from the recovered cells;

(c) infecting the cells by providing a solid support of a biocompatible substrate material being at least partially coated with a nanostructured TiO₂ film;

and culturing cells on the nanostructured TiO₂ film-coated support with a virion that carries genetic information;

(d) and re-administering the infected cells to the patient.

18. The method of claim 16 wherein the virions are retroviruses, adenoviruses, adeno-associated viruses (AAV) and any other viruses that can be utilized as vectors for genetic manipulation of cells.

19. The method according to claim 16 wherein the virions are genetically modified.

17. A method for gene therapy in vivo comprising the steps of:

(a) providing particles or a device of a nanostructured TiO₂ film-coated biocompatible material;

(b) loading the particles or device with virions; and

(c) implanting the virion-loaded particles or device into a tissue of a patient.

20. A method for cell replacement therapy comprising the steps of:

(a) providing particles or a device of a nanostructured TiO₂ film-coated biocompatible material;

(b) loading the particles or device with cells to be replaced in a patient; and

(c) implanting the cell-loaded particles or device into a tissue of a patient.

21. The method of claim 20 wherein the cells are genetically modified.

22. A method for the production of a solid support, comprising the steps of:

fabricating the solid support from a biocompatible substrate material;

;

depositing a nanostructured TiO₂ film at least a portion of the substrate material by nanoparticle deposition from a gas-phase; and

contacting the surface of the nanostructured TiO₂ film with virions and/or cells.

23. The method of claim 22 wherein the nanoparticle deposition from the gas-phase is carried out by means of supersonic cluster beam deposition (SCBD) using a pulsed microplasma cluster source.

24. A method of virus-mediated gene delivery to cells, comprising the steps of:

(a) providing a solid support of a biocompatible substrate material being at least partially coated with a nanostructured TiO₂ film;

(b) contacting the nanostructured TiO₂film-coated support with gene delivering virions;

(c) contacting the virions adhered on the surface of the nanostructured TiO₂film-coated support with a cell preparation; and

(d) culturing the cells for a time sufficient for gene delivery to occur.