Efficient deposition of nano-sized particles onto cells at an air liquid interface

Patent metadata

Field	Value
Publication number	US-12234440-B2
Application number	US-202016912486-A
Country	US
Kind code	B2
Filing date	Jun 25, 2020
Priority date	Jun 25, 2019
Publication date	Feb 25, 2025
Grant date	Feb 25, 2025

Abstract

An apparatus and method for the delivery and deposition of particles air-liquid-interface (ALI) cell cultures includes a sample inlet coupled to a growth tube having interior walls that are wet. The growth tube is configured to operate at a first temperature along a first length of the tube and a second temperature along a second length positioned between the first length and a growth tube outlet. The apparatus also includes a nozzle plate having a plurality of nozzles. The exposure chamber adapted to hold cell cultures at an air-liquid interface positioned underneath the plurality of nozzles and a temperature regulator adapted to control a temperature of the exposure chamber. The apparatus also includes a controller including instructions operable to cause the controller to maintain a relative humidity within the exposure chamber by controlling at least the second temperature of the growth tube and the temperature of the exposure chamber.

First claim

We claim:

1. An apparatus, comprising:
a sample inlet coupled to a plurality of laminar flow, condensational growth tubes, the plurality comprising a number of growth tubes, each of the plurality of growth tubes having interior walls that are wet such that a sample flow entering the inlet is split between the plurality of growth tubes, each condensational growth tube configured to operate at a first temperature along a first length of the tube which is adjacent to the inlet and a second temperature along a second length positioned between the first length and a growth tube outlet, the tube in the second length comprising a supersaturation region within the condensation growth tube;
a nozzle plate having a plurality of nozzles, the plurality of nozzles comprising a number of nozzles, each of the nozzles having an input configured to receive the output of one of the plurality of growth tubes and an output in an exposure chamber, the input having a greater area than the output such that the output focuses enlarged particles to hold in vitro cell culture systems positioned at a distance from the output, each nozzle having a nozzle diameter; and
a temperature regulator adapted to control a temperature of the nozzle plate and exposure chamber; and
a controller including instructions operable to cause the controller to maintain a relative humidity within the exposure chamber by controlling at least the second temperature of the growth tube and the temperature of the exposure chamber, and controlling a flow rate of CO2 introduced into the apparatus;
wherein the first length, the second length, the tube diameter, the number of the plurality of growth tubes, the number of nozzles, and the nozzle diameter of each of the plurality nozzles are configured to produce an output velocity of a flow at the output of less than 22 m/s for a sampling flow rate of 6.0 L/m and a deposition efficiency at inertial impaction velocities having a Stokes number above 0.2.

2. The apparatus of claim 1 wherein the nozzle plate includes one or more nozzles for each of the plurality of growth tubes, each nozzle having a converging flow guiding section between the input and the output exiting into the exposure chamber, the flow guiding section reducing a cross-sectional area of the incoming flow path.

3. The apparatus of claim 1 further including a second inlet coupled to a CO 2 source and provided adjacent to the sample inlet such that a sample flow provided to the sample inlet is mixed with CO 2 from the second inlet.

4. The apparatus of claim 1 wherein the condensational growth tube includes a third length positioned between the second length and the output, the third stage configured to have a third temperature, the third temperature adapted to be higher or lower than the second temperature and adapted to remove excess water vapor content in a supersaturated flow.

5. The apparatus of claim 4 further including a flow guiding adapter positioned between the third length of the growth tube and the nozzle plate, the flow guiding adapter including at least one channel positioned between the output of the condensational growth tube and the inputs of the nozzles, the channel being straight or angled relative to an axis passing through a center of the growth tube to mate with the input of the one or more nozzles.

6. The apparatus of claim 1 wherein the temperature regulator is configured to maintain the exposure chamber at a temperature which maintains cell viability based on a region of the respiratory system from which the cell originates.

7. The apparatus of claim 6 wherein the wherein the temperature regulator is configured to maintain the exposure chamber at about 37° C. lung cell viability or other temperatures for cells at other regions of the respiratory system.

8. The apparatus of claim 1 further including an exposure plate in the exposure chamber configured to mount one or more cell substrates for simultaneous exposure.

9. The apparatus of claim 8 wherein the exposure plate is rotatable and including a rotation motor.

10. An apparatus, comprising:
a sample inlet coupled to a plurality of growth tubes such that a sample flow provided to the inlet is split between the plurality of growth tubes, each growth tube having interior walls that are wet and configured to operate at a first temperature along a first length of the tube which is adjacent to the inlet and a second temperature along a second length positioned between the first length and a growth tube outlet;
a nozzle plate having a plurality of nozzles, at least one of the plurality of nozzles associated with each growth tube, each of the plurality of nozzles having an input configured to receive the output of one of the plurality of growth tubes and an output in an exposure chamber, the exposure chamber adapted to hold cell cultures at an air-liquid interface in a position below the plurality of nozzles, each nozzle having a nozzle diameter, and
a temperature regulator adapted to control a temperature of the exposure chamber; and
a controller including instructions operable to cause the controller to maintain a relative humidity within the exposure chamber by controlling at least the second temperature of the growth tube and the temperature of the exposure chamber, and controlling a flow rate of sampling flow introduced into the apparatus;
wherein the first length, the second length, the tube diameter and the nozzle diameter of each of the one or more nozzles are constructed to produce an output velocity of a flow at the output of less than 22 m/s for a sampling flow rate of 6.0 L/m.

11. An apparatus, comprising:
a sample inlet coupled to a plurality of growth tubes such that a sample flow provided to the inlet is split between the plurality of growth tubes, the plurality comprising a number of growth tubes, each of the growth tubes having interior walls that are wet, each growth tube configured to operate at a first temperature along a first length of the tube which is adjacent to the inlet and a second temperature along a second length positioned between the first length and a growth tube outlet;
a nozzle plate having a plurality of nozzles, the plurality of nozzles comprising a number of nozzles, each of the plurality of nozzles having an input configured to receive the output of one of the plurality of growth tubes and an output in an exposure chamber, the exposure chamber adapted to hold cell cultures at an air-liquid interface positioned underneath the plurality of nozzles, each of the plurality nozzles has a nozzle diameter;
a temperature regulator adapted to control a temperature of the exposure chamber; and
a controller including instructions operable to cause the controller to maintain a relative humidity within the exposure chamber by controlling at least the second temperature of the growth tube and the temperature of the exposure chamber, and controlling a flow rate of sampling flow introduced into the apparatus;
wherein the first length, the second length, the tube diameter, the number of the plurality of growth tubes, the number of nozzles, and the nozzle diameter of each of the plurality of nozzles are constructed to produce an output velocity of a flow at the output of less than 22 m/s for a sampling flow rate of 6.0 L/m and a deposition efficiency at inertial impaction velocities having a Stokes number above 0.2; and
wherein the exposure plate is rotatable and including a rotation motor.

Assignees

• Aerosol Dynamics Inc
• Univ Florida

Inventors

• Eiguren Fernandez Arantzazu
• Lewis Gregory Stephen
• Spielman Steven Russel
• Wu Chang-Yu
• Tilly Trevor Benson

Classifications

• C12N5/0062

  General methods for three-dimensional culture · CPC title

• C12N2513/00

  3D culture · CPC title

• C12M41/26

  of pH · CPC title

• C12N5/0006

  Modification of the membrane of cells, e.g. cell decoration · CPC title

• C12N2500/60

  Buffer, e.g. pH regulation, osmotic pressure · CPC title