I\'m still considering an oral Leflunomide try

Stem cell; cell based hair therapy
21

» Because of all the details in the patent concerning this topic, timing
» seems to be relavent in terms of when to apply the drugs.

The question could be phrased in a different way: why is there a wait period at all before applying the drugs?

I have a lot of thinking to do, but I am probably going to err on the side of starting the drugs a little early.

22

Im going to cut-and-paste and hypothesize thereafter:

Reepithelialization

In one aspect of this invention, the compositions of the invention are administered to a subject’s skin (examples of the skin location are the head, for example, the scalp, the eyebrow, or a scarred region) while the skin is in a state of reepithelialization. Reepithelialization is the process that occurs during formation of a new epidermis and can be characterized for the purposes of this invention by the lack of hair follicle morphogenesis (e.g., if within the tissue some cells are in the pre-placode stage of hair follicle formation), an embryonic-like state, in which the follicle regenerates, or by lack of a stratum corneum.

State of Reepithelialization Reepithelialization can be detected through inspection of the new epidermis where covering of the wound area by keratinocytes indicates reepithelialization. The presence of a keratinocytes can be seen with the naked eye as a white, glossy, shiny surface that gradually covers the open wound. Using a confocal microscope, keratinocytes can be visualized as a sheet of “cobblestone” looking cells. Reepithelialization can also be detected through the measurement of trans epidermal water loss (TEWL). TEWL decreases when the epithelial barrier is restored. Confocal scanning laser microscopy and/or optical coherence tomography can also be used to detect the state of

reepithelialization, where the presence of keratinocytes indicates reepithelialization.

The presence of a stratum corneum can be determined though visual inspection, direct observation of papillary blood vessels using a capillary microscope, or through a colorimetric redox reaction of a compound that reacts in the presence of live cells. For example, 0.01% nitrazine yellow applied to the skin will remain yellow if a stratum corneum is present, and will turn greenish brown if not. In another example 0.01% bromcresol purple applied to the skin will stay yellow if the stratum corneum is present and will turn purple if the stratum corneum is not present.

The area of reepithelialization can be, for example, between 0-2 millimeteres (mm) in width (e.g., 1 mm, 2 mm, 3 mπij or greater), 0-2 centimeters (cm) in width (e.g., 1 cm, 1.5 cm, and 2.0 cm) or greater. Optionally, the area of reepithelialization can be interfollicular. In some aspects of the invention, it is desirable to administer the compounds of the invention at a particular phase of reepithelialization. Stages at which compounds of the invention may preferably be administered and/or activated include periods:

■ after completion of the reepithelialization process (e.g., 3-12 days, or 9- 11 days after having disrupted the skin),
■ after or during the establishment of a stem cell population that will develop into a regenerated hair follicle (Ito et al, Nature 447, 316-320, May 2007),

Alternatively the compounds of the invention can be administered prior to epidermal disruption. In such embodiments, the compound may be formulated for controlled release such that the therapeutically active compound is released during reepithelialization or during a particular phase of …

DEPTH

The disruption of the epidermis can be induced between 3-12 days (e.g., 4-12, 5-12, 4-11, 6-11, 6-10, 6-9, 7-8, 5-11, 5-10, or 7-10 days) prior to the addition of the compositions of the invention.

Any of the above-described methods may be used to remove a precise amount of epidermal tissue. For example, the methods of abrasion described herein may be used to achieve:

• Removal of the stratum comeum through removal of the first 10-30 μm of dead skin cells.

• Removal of the stratum corneum and part or all of the epidermis by removing the first 30-100 μm of the skin. This is not deep enough to remove the sebaceous gland, bulge, or hair papilla of existing follicle structures.

MOST IMPORTANT VERBIAGE IN PATENT:

To determine whether human skin responded to EDIHN as did mouse skin, human skin was grafted onto SCID (immuno-defϊcient) mice and subjected to depilation by plucking and wound induction three days later. Seven days following wound induction, formation of new HF was observed in the human skin (Figure 2 IA; arrows indicate new HF) by hematoxylin and eosin staining of paraffin embedded tissue sections.

In additional experiments, adult human skin was grafted onto mice, abraded, and examined at 7 days post-abrasion. New HF were generated in the human skin, which mimicked normal hair follicle formation during fetal development, as evidenced by staining for S100A6 or S100A4 (Figure 21B).

23

» Im going to cut-and-paste and hypothesize thereafter:
»
»
»My comments are that the patent seems to indicate that if the compounds are put on while re-epilithialization is still going on, it must be OK because it notes “while” the skin is in a state of reepilithialization twice.
They then repeat the 3-12 days verbiage as a time that the compounds can be started over and over.

The wound depth is mentioned as less than 100 picometers, which aint much. This should not bring blood----it shouldn’t even effect sebaceous glands or even vellus hairs. If we are bleeding, we went too deep (my opinion).

That last paragraph is what has me most excited. “IN ADDITIONAL EXPERIMENTS” means so much to me. It wasn’t a one-time fluke. They got some de noveau hair in repeated experiment(s). That is plural. So we know at least TWICE more they got hair after dermabrasion. They didn’t even mention the plucking hair to prime the pump three days beforehand either. The hair germs were seen at seven days----which strongly leads me to believe that one shouldn’t wait longer than six-days post wounding to start with the EGF-inhibitors. I’ll probably go on day three----maybe even day two.

If TAGOHL or CAL notice anything Im concluding wrong, please let me know. I want us all to have big success. :ok:

24

» » Im going to cut-and-paste and hypothesize thereafter:
» »
» »
» »My comments are that the patent seems to indicate that if the compounds
» are put on while re-epilithialization is still going on, it must be OK
» because it notes “while” the skin is in a state of reepilithialization
» twice.
» They then repeat the 3-12 days verbiage as a time that the compounds can
» be started over and over.
»
» The wound depth is mentioned as less than 100 picometers, which aint much.
» This should not bring blood----it shouldn’t even effect sebaceous glands or
» even vellus hairs. If we are bleeding, we went too deep (my opinion).
»
» That last paragraph is what has me most excited. “IN
» ADDITIONAL EXPERIMENTS” means so much to me. It wasn’t a
» one-time fluke. They got some de noveau hair in repeated experiment(s).
» That is plural. So we know at least TWICE more they got hair after
» dermabrasion. They didn’t even mention the plucking hair to prime the pump
» three days beforehand either. The hair germs were seen at seven
» days----which strongly leads me to believe that one shouldn’t wait longer
» than six-days post wounding to start with the EGF-inhibitors. I’ll probably
» go on day three----maybe even day two.
»
»
»
»
» If TAGOHL or CAL notice anything Im concluding wrong, please let me know.
» I want us all to have big success. :ok:

Ok, can somebody tell me how may layers of skin is 100 picometers? Would a tca peel of 20 % do? I know the 1st 2 layers are just dead skin, a tca peel of about 20% should get you to the 3-5 layers I assume since it is rated as a medium peel.

25

Lately Ive been having computer problems. I have a bad driver that I know of, but other things also. This computer is from the early 00’s. It may be time to buy another one. My computer shut down six times this morning when I tried to get online—so that I had to cut the power to even get it back on, etc… Its telling me that I have serious errors and all that jazz… So if I dissapear from being online for a week or two, its not because Ive lost interest in abrasion, etc.

If I get the geftinib in the mail, I’ll be trying that experiment pretty soon therafter though…

26

It basically sounds to me like the wounding answer is:

Wait until the skin has reddened/peeled enough that it’s covered with a shiny layer.

That’s usually somewhere between two days and a week from what I recall about previous injuries in my life.