# US 11,433,231 B2 · Bioelectric stimulation for sonic hedgehog expression > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,433,231 B2 - Document ID: US11433231B2 - Status: Granted - Kind: B2 - Application number: 16/847,351 - Filed: 2020-04-13 - Published: 2020-10-15 - Granted: 2022-09-06 - Earliest claimed: 2019-04-15 - Assignees: Leonhardt Ventures LLC - Applicants: Leonhardt Ventures LLC - Inventors: Howard J. Leonhardt - Web page: https://patentgrep.com/patent/US11433231B2 ## CPC - A61N 1/0472: {Structure-related aspects} - A61N 1/36034: {specified by the stimulation parameters} - A61N 1/0492: {Patch electrodes (A61N1/0412, A61N1/0428 take precedence)} - A61N 1/205: {for promoting a biological process} - A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B) ## Abstract Described is a low voltage, pulsed electrical stimulation device for controlling expression of sonic hedgehog (“Shh”), a useful protein, by tissues. Also described are methods of enhancing expression of sonic hedgehog in cells, particularly a method of stimulating the expression and/or release of Shh in a cell having a gene encoding Shh, wherein the method includes applying a bioelectric signal of less than 50 Hz (e.g., 5 Hz, 10 Hz, or 20 Hz) at a pulse width duration of, e.g., 1 ms, to the cell (e.g., directly, indirectly, or wirelessly), and wherein the amount of Shh expression enhanced by this bioelectric signal is greater than that seen with a prior art bioelectric muscle stimulation or bioelectric muscle contraction alone as may be determined by, e.g., by an analysis of the upregulation of mRNA level/GAPDH fold gene expression in the cell. ## Claims ### Claim 1 A method for upregulating expression of Sonic hedgehog polypeptide in a tissue of a subject, the tissue selected from the group consisting of muscle, heart, liver, pancreas, skin, and lung, the method comprising: utilizing a bioelectric stimulator to apply a bioelectric signal to the tissue that upregulates the expression of Sonic hedgehog polypeptide in the tissue, wherein the bioelectric signal that upregulates expression of Sonic hedgehog in the tissue comprises: a biphasic pulse with a frequency range of 50 Hz to 100 Hz at a current greater than or equal to 100 μA at the tissue. ### Claim 2 A method of using a bioelectric stimulator to stimulate tissue of a subject, wherein the bioelectric stimulator is programmed to produce a bioelectric signal that upregulates expression of Sonic hedgehog in the tissue, the bioelectric signal comprising: a biphasic pulse with a frequency range of 50 Hz to 100 Hz at a current greater than or equal to 100 μA at the tissue, and wherein the subject is in need of wound healing, hair regeneration, and/or treatment of erectile dysfunction, the method comprising: connecting the bioelectric stimulator to the target tissue of the subject, and actuating the bioelectric stimulator to produce the programmed bioelectric signal so as to upregulate expression of Sonic hedgehog in the tissue. ### Claim 3 (depends on 2) The method according to claim 2, wherein the bioelectric signal provides a current of from 100 μA to 500 μA as measured three (3) mm deep into the tissue. ### Claim 4 (depends on 2) The method according to claim 2, wherein the bioelectric signal has a frequency/pulse width duration of 50 Hz/1 ms. ### Claim 5 (depends on 2) The method according to claim 2, wherein the bioelectric signal has a frequency/pulse width duration of 100 Hz/1 ms. ### Claim 6 A method of stimulating the expression of sonic hedgehog (“Shh”) in a cell having a gene encoding Shh, wherein the method comprises: applying to the cell a bioelectric signal of 5 Hz, 10 Hz, or 20 Hz at an adequate pulse width duration, wherein the amount of Shh expression enhanced by this bioelectric signal is greater than that seen with a generic bioelectric cell stimulation alone as may be determined by an analysis of the upregulation of mRNA level/GAPDH fold gene expression in the cell in each situation. ### Claim 7 (depends on 6) The method according to claim 6, wherein the bioelectric signal has a frequency/pulse width duration of 5 Hz/1 ms. ### Claim 8 (depends on 6) The method according to claim 6, wherein the bioelectric signal has a frequency/pulse width duration of 10 Hz/1 ms. ### Claim 9 (depends on 6) The method according to claim 6, wherein the bioelectric signal has a frequency/pulse width duration of 20 Hz/1 ms. ### Claim 10 A bioelectric stimulator programmed to produce at least one bioelectric signal that upregulates expression of Sonic hedgehog polypeptide in a target tissue, wherein the bioelectric signal is selected from the group consisting of a biphasic pulse at a frequency of 5 Hz with a pulse width duration of 1 ms at a current greater than or equal to 100 pA when measured three (3) mm deep into the target tissue, a biphasic pulse at a frequency of 10 Hz with a pulse width duration of 1 ms at a current greater than or equal to 100 pA when measured three (3) mm deep into the target tissue, a biphasic pulse at a frequency of 20 Hz with a pulse width duration of 1 ms at a current greater than or equal to 100 pA when measured three (3) mm deep into the target tissue, and a biphasic pulse at a frequency of 40 Hz with a pulse width duration of 1 ms at a current greater than or equal to 100 pA when measured three (3) mm deep into the target tissue. ### Claim 11 (depends on 10) The bioelectric stimulator of claim 10, wherein the bioelectric signal has a frequency of 5 Hz with a pulse width duration of 1 ms. ### Claim 12 (depends on 10) The bioelectric stimulator of claim 10, wherein the bioelectric signal has a frequency of 20 Hz with a pulse width duration of 1 ms. ### Claim 13 (depends on 10) The bioelectric stimulator of claim 10, wherein the bioelectric signal has a frequency of 40 Hz with a pulse width duration of 1 ms. ### Claim 14 (depends on 10) A method of using the bioelectric stimulator of claim 10 to stimulate tissue of a subject, wherein the subject is in need of wound healing, hair regeneration, and/or treatment of erectile dysfunction, the method comprising: connecting the bioelectric stimulator to tissue of the subject, and actuating the bioelectric stimulator to produce the programmed bioelectric signal so as to upregulate expression of Sonic hedgehog in the tissue. ### Claim 15 (depends on 14) The method according to claim 14, wherein the bioelectric signal provides a current of from 100 μA to 500 μA as measured three (3) mm deep into the tissue. ### Claim 16 (depends on 14) The method according to claim 14, wherein the bioelectric signal has a frequency/pulse width duration of 5 Hz/1 ms.