Calcium controlled activation of platelets via electrical stimulation

Patent metadata

Field	Value
Publication number	US-2017191052-A1
Application number	US-201514984988-A
Country	US
Kind code	A1
Filing date	Dec 30, 2015
Priority date	Dec 30, 2015
Publication date	Jul 6, 2017
Grant date	—

Abstract

The present disclosure relates to the generation of an activated platelet product in which one or more of the presence or absence of clots, the timing of clot formation (if present), and/or the mechanical strength of clots (if present) is controlled by the presence or concentration of calcium ions during the activation process. In certain embodiments, the calcium ion concentration is controlled in the presence of pulsed electric fields or a chemical activator (e.g., thrombin) as part of the activation process.

First claim

1 . A method for generating an activated product comprising:
preparing a platelet-rich plasma (PRP) sample for activation, wherein the PRP sample is prepared by adding calcium ions at a respective concentration, wherein the respective concentration is selected based upon whether clots are to be present in an activated product composition generated using the PRP sample and, if clotting is to be present, one or more of a time until clot formation or a mechanical strength of the clots; positioning the PRP sample with respect to electrodes of an electromagnetic stimulation apparatus; specifying a set of electrical pulse parameters; and exposing the PRP sample to one or more electrical pulses generated in accordance with the parameter values, wherein the PRP sample, when exposed to the one or more electrical pulses, yields an activated product composition comprising one or more growth factors and having the specified clotting characteristics.

2 . The method of claim 1 , wherein the set of electrical pulse parameters is selected based at least upon in part upon the selected concentration of calcium ions.

3 . The method of claim 1 , wherein the same electrical pulse parameters but different concentrations of calcium ions yield different clotting characteristics in the activated product composition.

4 . The method of claim 1 , wherein the sample comprises one of a platelet rich plasma sample, a platelet suspension, or a whole blood sample.

5 . The method of claim 1 , wherein the relative levels of the one or more growth factors are determined by one or both of the set of electrical pulse parameters or the added calcium ions.

6 . The method of claim 1 , wherein positioning the sample between the electrodes comprises flowing the sample through a conduit between the electrodes, wherein the activated product composition has relative levels of the one or more factors determined at least in part by one or both of the conduit diameter or the flow rate of the sample through the conduit.

7 . The method of claim 1 , comprising adding CaCl 2 to anticoagulant-treated PRP sample in a concentration in the range of about 2.5 mM to about 20 mM.

8 . The method of claim 1 , wherein the concentration of calcium ions is selected from the plurality of possible concentrations corresponding to 2.5 mM 5.0 mM, 7.5 mM, 10 mM, 15 mM, 20 mM, or 25 mM CaCl 2 being added to the anticoagulant-treated PRP sample.

9 . The method of claim 1 , wherein a first calcium concentration that is less than a second calcium ion concentration results in an initial clot formation that is slower than that observed at the second calcium ion concentration.

10 . The method of claim 1 , wherein a first calcium concentration that is less than a second calcium ion concentration results in a clot mechanical strength that is less than that observed at the second calcium ion concentration.

11 . A method for generating an activated product comprising:
preparing an anticoagulant-treated platelet-rich plasma (PRP) sample for activation; adding calcium ions to the PRP sample to achieve a calcium ion concentration selected from a range of possible concentrations, wherein the concentration is selected based upon target levels of one or more growth factors to be present in an activated product composition generated using the PRP sample; and exposing the PRP sample to electrical activation stimulus, wherein the PRP sample, when exposed to the electrical activation stimulus, yields an activated product composition comprising the one or more growth factors at the target levels, wherein varying the calcium ion concentration without varying the electrical activation stimulus changes one or both of the absolute or relative levels of the one or more growth factors.

12 . The method of claim 11 , wherein one or more dotting characteristics are also determined by the calcium ion concentration.

13 . The method of claim 11 , wherein adding calcium ions to the PRP sample comprises adding CaCl 2 to the PRP sample in a concentration in the range of about 2.5 mM to about 20 mM.

14 . The method of claim 11 , wherein the concentration of calcium ions is selected from a plurality of possible concentrations corresponding to 2.5 mM 5.0 mM, 7.5 mM, 10 mM, 15 mM, 20 mM, or 25 mM CaCl 2 being added to the PRP sample.

15 . The method of claim 11 , wherein a first calcium concentration that is less than a second calcium ion concentration results in an initial clot formation that is quicker than that observed at the second calcium ion concentration.

16 . The method of claim 11 , wherein a first calcium concentration that is less than a second calcium ion concentration results in a clot mechanical strength that is less than that observed at the second calcium ion concentration.

17 . A method for controlling clot mechanical strength in a platelet gel, comprising:
determining a prospective mechanical strength of one or more clots to be generated in the platelet gel, wherein the prospective mechanical strength is greater than what would be observed by generating the platelet gel using thrombin alone; based on the prospective mechanical strength, selecting a calcium ion concentration corresponding to the prospective mechanical strength from among a plurality of calcium ion concentrations; generating the platelet gel by activating a platelet-rich plasma (PRP) sample comprising calcium ions at the selected calcium ion concentration, wherein the PRP sample is activated using electrical stimulus, and wherein the platelet gel comprises clots that, once formed, have the prospective mechanical strength.

18 . The method of claim 17 , wherein, for the plurality of calcium ion concentrations, higher calcium ion concentrations correspond to greater mechanical strength of the clots.

19 . The method of claim 17 , wherein the plurality of calcium ion concentrations is within a range corresponding to what is generated by addition of about 2.5 mM to about 20 mM CaCl 2 to the PRP sample.

20 . The method of claim 17 , wherein the calcium ion concentration is selected from the plurality of possible concentrations corresponding to 2.5 mM 5.0 mM, 7.5 mM, 10 mM, 15 mM, 20 mM, or 25 mM CaCl 2 being added to the PRP sample.

Assignees

• Gen Electric

Inventors

• Neculaes Vasile Bogdan
• Torres Andrew Soliz
• Klopman Steve Lambert

Classifications

• A61P17/02

  for treating wounds, ulcers, burns, scars, keloids, or the like · CPC title

• A61K33/06

  Aluminium, calcium or magnesium; Compounds thereof {, e.g. clay} · CPC title

• C12N13/00 Primary

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12N5/0644

  Platelets; Megakaryocytes · CPC title

• A61K38/18 Primary

  Growth factors; Growth regulators · CPC title