Hypothermic 3d bioprinting of living tissues supported by perfusable vasculature

Patent metadata

Field	Value
Publication number	US-2018002658-A1
Application number	US-201715709392-A
Country	US
Kind code	A1
Filing date	Sep 19, 2017
Priority date	Mar 20, 2015
Publication date	Jan 4, 2018
Grant date	—

Abstract

The present disclosure provides compositions and methods for producing hydrogel matrix constructs. Methods of using hydrogel matrix constructs for tissue repair and regeneration and for the oxygenation of red blood cells are also disclosed.

First claim

What is claimed is:

1 . A prepolymerization solution comprising: a photosensitive polymer having a molecular weight of greater than 2,000 Daltons and comprising at least two vinyl groups per molecule of polymer; a photoinitiator; and a biocompatible, light-absorbing additive material suitable to control light penetration.

2 . The prepolymerization solution of claim 1 further comprising a cell.

3 . The prepolymerization solution of claim 1 , wherein the biocompatible, light-absorbing additive material is organic.

4 . The prepolymerization solution of claim 1 , wherein the biocompatible, light-absorbing additive material is tartrazine or curcumin.

5 . The prepolymerization solution of claim 1 comprising a water content of 10 wt % to about 99.5 wt %.

6 . The prepolymerization solution of claim 1 , wherein the polymer is poly(ethylene glycol) and wherein the at least two vinyl groups are methacrylate or acrylamide.

7 . The prepolymerization solution of claim 1 wherein the photo-initiator is lithium acylphosphinate.

8 . A composition comprising:
a hydrogel matrix comprising a plurality of layers, each layer comprising a cross-linked polymer network formed from a photosensitive polymer having a molecular weight greater than 2,000 Daltons, wherein the hydrogel matrix has a elastic modulus of from about 1 kilopascal to about 200 kilopascals; a first elongated void in the hydrogel matrix providing a first tubular channel; a second elongated void in the hydrogel matrix providing a second tubular channel; wherein the first tubular channel and the second tubular channel are perfusable; wherein the first tubular channel does not intersect the second tubular channel; and wherein the second tubular channel interpenetrates the first tubular channel.

9 . The composition of claim 8 , wherein the first tubular channel or second tubular channel is lined with cells.

10 . The composition of claim 8 , wherein the first and second tubular channel each comprise a horizontal segment, wherein a cross-section in the horizontal segment intersects more than one layer of the bulk hydrogel matrix.

11 . The composition of claim 8 , wherein the hydrogel matrix is porous.

12 . The composition of claim 8 , wherein the first tubular channel is filled with a fluid selected from the group consisting of culture media, blood, urine, and bile.

13 . The composition of claim 8 , wherein the second tubular channel is filled with oxygen.

14 . The composition of claim 8 , wherein the first tubular channel and second tubular channel have a diameter of 500 microns or less.

15 . The composition of claim 8 , wherein each layer of the hydrogel matrix has a thickness of 100 microns or less.

16 . The composition of claim 8 , wherein the first tubular channel and second tubular channel are expandable in response to increases in pressure therein.

17 . The composition of claim 8 , wherein one or more layers of the hydrogel matrix has cells embedded therein.

18 . A process for manufacturing a multi-layer hydrogel matrix construct, comprising:
a) creating a 3D model of the multi-layer hydrogel matrix construct using a design software, wherein the 3D model of the multi-layer hydrogel matrix construct comprises a first computational algorithm that yields a first elongated void in the multi-layer hydrogel matrix construct providing a first tubular channel, and a second computational algorithm that yields a second elongated void in the multi-layer hydrogel matrix construct providing a second tubular channel, wherein the second computational algorithm results in the second tubular channel interpenetrating the first tubular channel; b) converting the 3D model to a format suitable for use in a 3D printing software to yield a formatted 3D model; c) importing the formatted 3D model into the 3D printing software, wherein the 3D printing software is programmed to slice the formatted 3D model into multiple 2D xy images; d) supplying a prepolymerization solution to a vat associated with a build platform of a 3D printer, wherein the vat is transparent, and wherein the prepolymerization solution comprises a photosensitive polymer having a molecular weight of greater than 2,000 Daltons and at least two vinyl groups per molecule of polymer, a light-absorbing additive material to control light penetration, and a photoinitiator; e) positioning a mobile Z-axis stage of the 3D printer at a distance from the vat, wherein the Z-axis stage includes a surface sufficient for gelled material to adhere thereto, wherein the distance between the surface and an inner bottom surface of the vat is equivalent to a desired layer thickness of the multi-layer hydrogel matrix construct; f) projecting a pattern being onto the inner bottom surface of the vat, wherein the pattern corresponds to one of the multiple 2D xy images; g) polymerizing a layer of the multi-layer hydrogel matrix construct consistent with the pattern; and h) repeating steps d through g one or more times, wherein the mobile Z-axis stage is moved so that the distance moved is equivalent to the desired thickness of each subsequent layer, and wherein the same or a different pattern is displayed for each subsequent layer.

19 . The process of claim 18 , wherein the first computational algorithm is a Hilbert Curve, derived from knot theory, or conforms to Murray's law.

20 . The process of claim 18 , wherein the prepolymerization solution further comprises one or more cells.

Assignees

• Univ Rice William M

Inventors

• Miller Jordan
• Ta Anderson
• Grigoryan Bagrat

Classifications

• B29C64/393

  for controlling or regulating additive manufacturing processes · CPC title

• B33Y10/00

  Processes of additive manufacturing · CPC title

• B29K2071/02

  Polyalkylene oxides, e.g. PEO, i.e. polyethylene oxide, or derivatives thereof · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• B33Y70/00

  Materials specially adapted for additive manufacturing · CPC title