Understanding Mesenchymal to Epithelial cell transition may be key for neo-growth plates

This study is huge because we can get adult epithelial cells to potentially form new growth plates.  Since the resting zone is composed of mainly endothelial-like cells and the resting zone is the foundation of the growth plate.  Understanding how to transition adult mesenchymal stem cells into endothelial cells may be the basis for forming a new growth plate.

Epithelial-mesenchymal transition and mesenchymal-epithelial transition response during differentiation of growth-plate chondrocytes in endochondral ossification.

“For linear longitudinal bone elongation, the stem-like progenitor chondrocytes distributed in resting zone (RZ) of growth plate have a capacity to differentiate towards the spindle chondrocytes in proliferative zone (PZ), then towards the columnar and tightly adjacent chondrocytes in hypertrophic zone (HZ). We hypothesized this process of endochondral ossification with cells morphological change was occurred along with the inter-conversion between epithelial to mesenchymal cell types. Consistent with this hypothesis, our study demonstrated the chondrocytes highly expressed mesenchymal-like biomarkers and loss of epithelial surface markers in PZ, while converse in RZ and HZ of the growth plate in mice distal tibia in vivo. To further determine these process and correlation regulatory pathway, the 4-week old male and female mice were treated with estradiol cypionate or oxandrolone, then investigated the response of epithelial- and mesenchymal biomarkers, and demonstrated that estrogen blocked the EMT process from RZ to PZ while androgen promoted MET from PZ to HZ. Our observations supported the hypotheses that the growth plate firstly go through EMT from RZ to PZ, then MET process from PZ to HZ during the epiphyseal fusion. Our results could interpret the different roles of estrogen and androgen in growth plate cartilage when endochondral ossification.”

In Epiphelial to Mesenchymal Cell Transition, cells lose cell-cell adhesion and cell polarity properties to become more migratory mesenchymal cells.  Since the growth plate firstly grows through EMT at the RZ to PZ that is the most important stage as that causes the formation of the growth plate.  However, the cell condensation stage to establish the resting cell zone should first require epithelial cell types as cell to cell adhesion would be needed.

“Multiple tissues differentiation and organs formation in embryonic development arise from a
series of conversion from epithelial to mesenchymal cells, through epithelial to mesenchymal
transition (EMT) or mesenchymal to epithelial transition (MET). In primary EMT process,
the primitive epithelia lose their characterization of rounded shape, sequential arrangement
and compact junctions to convert a population of spindle, loosely organized but motile mesenchymal cells for gastrulation formation and neural crest migration. Then, after a transient
epithelial structure condensation through MET, these population in notochord, somites, somatopleure and splanchnopleure derived from mesoderm generate mesenchymal cells which have ability to differentiate into specific cells types of diverse tissues via the secondary EMT”

” the neural crest cells migrate to somites of mesoderm following stereotyped pathways and undergo a secondary EMT to generate mesenchymal condensation.  These mesenchymal cells differentiate into osteoprogenitors for intramembranous ossification and chondrocytes for endochondral ossification. ”

” Pluripotent stem cells exhibit epithelial characteristics, down-regulate the epithelial markers such as Cdh1, Cldn6, Epcam and enhance the mesenchymal markers including Snai1/2, Zeb1,

” estrogen administration maintains the epithelial type genes expression in growth plate particularly in RZ implies that estrogen appears to block EMT process.”

“Not like human or rabbit, the expression of estrogen receptors within HZ of growth plate in mice and rat was extremely low until at the last time point prior to epiphyseal fusion, which also
reflects the less effect of estradiol cypionate to in the HZ in our study. Conversely, Androgen
effectively promotes EMT for  chondrocytes differentiation”

” estrogen may interdict TGF-beta, then further repress Smad3 expression, so that
postpone chondrocytes differentiation via EMT blocking. ”

Epithelial cells may already exist in adult bone marrow.

Epithelial cells in bone marrow: do they matter?

“epithelial-like cells can be detected in the bone marrow of many patients not known to have cancer. ”

Inducing a mesenchymal to epithelial transition would be difficult as I haven’t found any studies of it occuring due to physiological stimuli.  Another possibility though would be to have a growth plate without a resting zone as the proliferative zone consists mainly of mesenchymal cells.  The viability of this depends on the viability of a growth plate without a resting zone.  The resting zone may play a role in growth plate orientation which makes sense as epithelial cells tend to be involved in cell polarity and cell adhesion.

Here’s a diagram of the mesenchymal-epithelial transition:

mesenchymal-epithelial transition

LSJL upregulates Pcdhb2(protocadherin beta 2), Cdh13, Ctnna3, Fat1(a cell adhesion model).  It downs regulates protocadherin subfamily A, 4(Pcdhga4), AK002616(a miscellaneous Cadherin related protein), Celsr2, Cdh15, Cdh11.  So LSJL has the definite potential to affect the mesenchymal-epithelial transition although how isn’t clear.

For other parts LSJL downregulates Cldn13, Dsp(Desmoplakin isoform 1, the downregulation of this gene suggests that LSJL likely encourages the epithelial to mesenchymal transition but that doesn’t mean that it doesn’t encourage the mesenchymal to epithelial transition as well), and upregulates a gene related to Cldn19, Muc3.  It also downregulates an anti-mucin gene.

This diagram mentions the reverse markers:

Here’s another diagram:
another EMT diagram

According to Actin stress fibres and cell-cell adhesion molecules in tendons: organisation in vivo and response to mechanical loading of tendon cells in vitro.

“Tendons consist of parallel longitudinal rows of cells separated by collagen fibres. The cells are in intimate contact longitudinally within rows, and laterally via sheet-like lateral cell processes between rows. At points of contact, they are linked by gap junctions. Since tendons stretch under load, such cell contacts require protection. Here we describe the organisation of the actin cytoskeleton and actin-based cell-cell interactions in vivo and examine the effect of cyclic tensile loading on tendon cells in vitro. Cells within longitudinal rows contained short longitudinally running actin stress fibres. Each fibre was aligned with similar fibres in the cells longitudinally on either side, and fibres appeared to be linked via adherens junctions. Overall, these formed long oriented rows of stress fibres running along the rows of tendon cells. In culture, junctional components n-cadherin{this increase is actually not good news for creation of epithelial cells as this is a mesenchymal marker} and vinculin and the stress fibre component tropomyosin increased in strained cultures, whereas actin levels remained constant. These results suggest that: (1) cells are linked via actin-associated adherens junctions along the line of principal strain; and (2) under load, cells appear to attach themselves more strongly together, and assemble more of their cytoplasmic actin into stress fibres with tropomyosin. Taken together, this suggests that cell-cell contacts are protected during stretch, and also that the stress fibres, which are contractile, may provide an active mechanism for recovery from stretch. In addition, stress fibres are ideally oriented to monitor tensile load and thus may be important in mechanotransduction and the generation of signals passed via the gap junction network.”

So according to this in response to load cells may establish more cell-cell contact characteristic of epithelial cells.

Update On Older Research – Melatonin and Growing Taller

When I first started writing on the website, I remember that people were talking about this supplement combination which was supposed to help you grow taller. M.E.N.S.

That was the acronym used for sleep, exercise, niacin, and melatonin. I wanted to talk about melatonin.

I recently started to take melatonin just to help me sleep and I remembered that I never really did much research on Melatonin. A quick Google Search on the link between melatonin and the possibility that it can help you grow taller reveals many of the older posts written maybe 4–7 years ago, derived from the now dead GrowTallForum.com website.

People believed in this supplement combination. Melatonin was promoted a lot by people of that old forum because the thinking was that since it helps you sleep, and growth hormones are primarily released when you are sleeping, melatonin would mean you have a greater chance of having growth hormones being pushed into your system.

I looked at the link. Is there something there?

I refer to 7 studies, some of them recently published just in 2014.

The 3rd study suggests that Melatonin does have this ability to turn MSCs into the chondrogenic lineage. Other sources (#7) says that for the growing child, over-expression of melatonin in the system can cause abnormal chondrocyte growth. It turns out that young kids who have idiopathic scoliosis actually have longer bones (vertebral column and arm length) than their peers. The curvature of the vertebral is because the anterior region of the vertebral is growing faster than the posterior region.

Out of all of the 7 studies, I felt that study #1 was the most insightful.

Short stature is most often caused by 2 ways.

  1. Idiopathic Short Stature (ISS)
  2. Growth Hormone Deficiency (GHD)

It seems that for children who suffer from GHD, they have GHD because of the overexpression of melatonin.

This means that based on this 1 studies, when we generalize the conclusion, it seems that having too much melatonin in the child’s system is bad for their height. However, this condition is something that can be medically treated, unlike ISS, which most family doctors would say makes the kids “genetically predisposed to be short later in life”.

So while MSCs might be helped slightly by Melatonin, too much Melatonin is a bad thing. Referring to study #5, the conclusion states the following for peri-pubertal children. – “These findings indicated that melatonin could inhibit the proliferation and stimulate differentiation of GPC (Growth Plate Chondrocytes) in human”{Tyler-Actually this could be a thing for a height growth as peak chondrocyte hypertrophy is more important than chondrocyte proliferation for peak height growth}

Taking Melatonin does not work. Try to figure out another type of supplement to take to sleep easier.

Excess Pituitary Stimulated Growth Hormones Increases Male Genitalia aka Penis Size

Something I read from the reddit/r/short threads recently made me remember something I discovered years ago. It seems that male genitalia, (aka Penis Size) can be altered/increased through excess levels of hormones.

I just don't know what the title might be, just please read it, not because of me.

Whoever the poster was, I am guessing that they are feeling the symptoms of a pituitary related tumor, whether benign or malignant. The tumor is stimulating excess HGH release into their system.

Like they said they are already 26, which suggest that their normal growth plates should be closed. This recent events which are indeed unusual for most people doesn’t seem to surprise me. I had guessed that excess HGH can cause even people with closed growth plates to gain 1 full inch in height, just based on the fact that there are still thin slivers of hyaline cartilage tissue in their vertebrate, tibia, femur, etc. which goes through hypertrophy. When you combine all of the hypertrophy the hyaline articular cartilage locations goes through, the total increase in height would often be around 1 inch.

As for height increase, this guy went from 6′ 2″ to 6′ 3″. His height will most likely not increase any further. However, he stated the other fact. His genitalia increased. When I think about this, it all starts to make sense.

Let’s go back a little.

For the normal, average, heterosexual, American men, the most common insecurity might not be their size, in terms of height/stature, but their size as in their “genitalia”. There are in fact entire internet forums dedicated by young men who actively do exercises to increase the size of their genitalia. Since the forums seem to be quite active, I am guessing that these guys who have that pursuit are slightly more successful in their endeavor than ours.

I had proposed in previous posts the idea that stereotypes have in them kernels of truth. I theorized that the stereotype that African American males have larger than average sized genitalia is true. The reason that is the truth is because I guessed that African Americans have higher than average levels of IGF-1 which goes through their system when they are going through puberty.

Higher levels of IGF-1 suggests higher rates of cardiovascular system related medical conditions later in life (diabetes, prostate cancer) which also has a slight correlation to having an earlier than average puberty. I am not sure, but I think I remember seeing studies which show that African American females (at least in urban locations) goes through puberty around 0.5 years earlier than their Caucasian American counterparts. This gives them a slightly less time to grow, assuming all other factors in their growth progression is the same.

Let’s remember our endocrine system. Pituitary gland derived GH goes into the liver, which converts it to IGF-1 which goes to the chondrocyte area (growth plate) of bones making them longer. We also learned that IGF-1 can be created located in the tissue right next to the growth plate. IGF-1 is created in both areas.

If my theory on the link between African Americans and increased IGF-1 levels during puberty is true, it would explain the increase in tissue (size and number) that is not just in bones.

This guy who posted stated that he noticed himself growing again. The obvious guess is that his pituitary gland has gone into overdrive, secreting GH, going to the lier, converting into IGF-1 and that is what is contributing to making his genitalia larger.

Now, this is not the only way chemically. There are a LOT of other options for men to increase the size of their genitalia. (think Relaxin, Prostaglandin ES , Prostaglandin Alpha F2.). In fact, it seems that taking something as simple as Niacin or Nitric Oxide (found in most GNC stores) would be able to do that.

Here is what most people need to realize about growing tissue in their body. Pituitary Gland derived HGH can grow almost all forms of tissue in the human body except for bones, at least interstitially.

If you have ever seen most people suffering from acromegaly, or maybe gigantism when they were younger, they often have wider than average noses, and have wider than average upper torso. The GH going through their system is expanding the costal cartilage in their sternum and the fibrocartilage in their nose even when they are in their 30s, and 40s, of course in small levels.

In fact, analogous to how men in today’s American society worry about the size of their genitalia, American females are too concerned with their mammary gland organs (breasts).

Too many American females get plastic surgery to make their breasts larger, to supposedly give themselves higher levels of confidence/self-esteem. What they don’t understand is that nature (being the generous “mother” that she is) figures out a way to give females who have smaller than average mammary glands larger ones, once they have gone through the process of pregnancy.

In fact, the actress Mila Kunis went on the Conan O’Brien show recently and talked about her experiences in adjusting to the fact that her mammary glands have increased from pregnancy, and now she needs to wear a bra, which she has not had to do throughout her adult life before. So girls, stop going to the plastic surgeon. Just wait for pregnancy to do it naturally. Sure, there is also the added side effect of increased nipple and aereola size, which is for the baby’s mouth. However, the mammary glands definitely increase in size.

Some people might counter and say that the increase is not permanent. I agree on that point, since based on female menstrual cycles, the breasts’ size increases and decreases based on the monthly cycle. However, pregnancy does cause certain chemicals to go often, which have such a dramatic affect on the female body that often the physiological and anatomical changes are completely permanent.

(I am referring to the fact that many caucasian females who had blonde hair before throughout their entire lives had their hair color changed to becoming brunettes because of the release of excess melanin from pregnancy. In fact, you will find many stories of females who had their hair progressively get darker and darker over time after each pregnancy they go through.)

From pregnancy, women gain these changes in their body

  • Feet size increase – Shoe sizes often go up by 1 full size, permanently. This is due to relaxin.
  • Breast size increase – I suspect at least 25% of all women who have gone through multiple pregnancy would notice an increase in cup/bra size that is permanent.
  • Hair color changes – Most pronounced in caucasian females. Light blonde hair turns progressively darker/more brown after each pregnancy.
  • Body hair changes – Increased levels of hair on arms/legs. Also, hair might sprout up in areas of body which wasn’t there before.
  • Hip size increase – Some medical profession might argue with me and say that certain women can never loss the extra 15 lbs they gain from their first pregnancy. They account hip size increase due to fat deposits that become an extra layer. However, I might argue against this and say that hip size increases can be due to realignment of the pelvic/sacroiliac girdle, causing the pubic symphysis to become stretched out.
  • Height increase – Now, this is the phenomena which is most rare. I had guessed that this only happens to maybe 1 our of every 10,000 women.

Interestingly, just today some other woman commented on a previous post “Another Case Of Pregnancy Causing Woman To Grow Taller And Increase In Height”.

Let me show you guys just how often I get messages to this website to that post about this phenomena.





Taking Dramamine Seems To Help Children and Adults Grow Taller

While I was doing chemical research for something else today, specifically on the possibility of starting my own chemistry based cosmetic company (similar to what Jessica Alba did with her Honest company to become a billionaire) I found myself looking into at the active ingredients of many of the most common items in one’s bathroom.

It took maybe 3 hours of checking, scouring, and reading (on Wikipedia) to figure out what exactly were the compounds found in my mouthwash, soap, toothpaste, hand lotion, and sunscreen.

Eventually I came across something that was lying on my bathroom medicine cabinet, Benadryl.

I once remembered a friend of mine telling me that you can knock yourself out by drinking a lot of Vick’s Nyquill (aka cough syrup), which has the active chemical compounds of the following below… (source)

Active ingredients (in each 15 ml TBSP) – Purpose

  • Acetaminophen 325 mg  – Pain reliever/fever reducer
  • Dextromethorphan HBr 15 mg – Cough suppressant
  • Doxylamine succinate 6.25 mg – Antihistamine

Antihistamines are a very common type of drug. So from Vick’s Nyquill, I wanted to know the active ingredient of Benadryl. Does Benadryl have a similar effect as Nyquill, where if you take too much, you will get knocked out?

(Side Fun Fact: Interestingly, people often mistake the idea of chloroform as a drug used to knock people out, but it is actually Ether. Chloroform is talked a lot about in Movies, and shown used in Kidnappings, but Ether would be more effective, since it is faster acting on the human system)

(Side Fun Fact #2: Similarly, there is more than one way to make truth serum, which really just lowers people’s inhibitions and acts as a sedative, making them groggy. The recent news about thieves who steal from tourists using the NIghtshade derived Scopolamine and Atropa belladonna. You can also use Sodium Pentothial (aka Sodium Pentathol.)

So what are the active ingredients in Benadryl, the Anti-Histamine Drug most often used to treat allergies like hay fever?

Well, Benadryl is actually a Brand Name for something else, called Diphenhydramine. In the UK, the active ingredient would be acrivastine and/or cetirizine.

While I was doing more research into the compound Diphenhydramine, It led me to an article on Meclizine, aka Meclozine. Medical professionals knew about the bad side effects of Diphenhydramine for a while, and wanted a type of anti-histamine which did not have that type of drowsiness which comes with it.

In my own searching, I remember writing about Meclozine as a one of the most biggest breakthroughs we have made on this website. Tyler back in 2014 wrote a post entitled Game Changing Breakthrough OTC Height Supplement-Meclozine. I myself also wrote about it entitled Achondroplasia Treatment and Increase Height Using Meclozine aka Meclizine – Great News For Parents!

The science behind it seems to be valid. However, there have been a few sources we found on the internet which suggests the exact opposite. Study of possible correlation between BODY HEIGHT DECREASED and MECLOZINE (MECLOZINE)

This one source however does not take away from the multiple sources which show that Meclizine has bone enhancing abilities.

Google patents – Therapeutic agent for systemic bone disease and use thereof – WO 2014141847 A1

This patent was based on an idea to treat achondroplasia by having as one of the active ingredients Meclizine, which they noted is OTC (Over-The-Counter). These same inventors from Japan, wrote a piece called “Meclozine, Motion Sickness Medicine, Has The Therapeutic Potential In Achondroplasia” – (Masaki Matsushita).

It is interesting that Meclozine (aka Dramamine) has sort of replaced the Benadryl/Diphenhydramine as the type of antihistamine to use now.

It seems that Meclozine is actually so common, and easy to get that you can buy it from Amazon. Before, Tyler Linked to the chemical. Here it is again, for sale from Amazon for around $35. (No affiliate link)

Someone asked Tyler whether taking the Meclozine would work with people with closed growth plates. Tyler said that you should take it with doing LSJL. I will let the readers decide on whether it will work for adults.

However, for young children, this chemical seems to have noticeable effects, especially if you get the compound into your blood, and not through your digestive system (aka orally).

Teriparatide potential height increase tool

Teriparatide Improves Trabecular Osteoporosis but Simultaneously Promotes Ankylosis of the Spine in the Twy Mouse Model for Diffuse Idiopathic Skeletal Hyperostosis.

Full study->teriparitadeboneformation

“Diffuse idiopathic skeletal hyperostosis (DISH) is a common skeletal disorder in the elderly, which can develop into periosteal hyperostosis and paradoxically into immobilization-associated trabecular osteoporosis. The bone anabolic agent, teriparatide (TPD), seems to be a rational treatment for the immobilization-associated osteoporosis. However, it can lead to development of hyperostosis lesions in DISH patients. Here, we demonstrate TPD effectively treats trabecular osteoporosis while simultaneously promoting ankylosis of the spine in DISH model tiptoe-walking Yoshimura (twy) mice, compared with the ICR mice. Eighteen male twy mice were divided into three groups, and ICR mice were used as a normal control. Subcutaneous injections of TPD or phosphate-buffered saline (PBS) were performed according to three dosing regimens; 40 µg/kg once daily (TPD × 1 group), 40 µg/kg three times daily (TPD × 3 group), and PBS (control; Ctl group). Treatment was commenced at the age of 7 weeks and continued for 5 weeks. Micro-computed tomography (µCT) and histological analysis were performed. Longitudinal µCT study revealed that trabecular bone volume in both the vertebral body and distal femur decreased with time in the Ctl group, but increased dramatically in the TPD × 3 group. The twy mice developed ankylosis of the spine, the progression of which was accelerated with TPD therapy. We also confirmed that TPD therapy promoted ossification of spinal ligaments. Histomorphometrical study revealed that TPD treatment increased bone formation at the vertebrae enthesis region{This is the key to where this has promise} and in the trabecular bone. TPD therapy effectively treats trabecular osteoporosis, but potentially promotes ankylosis of the spine in patients with DISH.”

“The twy mice are mutant mice showing multiple osteochondral lesions, and have been used as a model for DISH and ossification of the posterior longitudinal ligament”

Teriparatide increased ectopic calcification(Fig3C).

“both TPD × 1 and TPD × 3 treatments promoted ectopic calcification. Histologically, there were no cells inside the ectopic calcification but there were fibroblast- or osteoblast-like cells around the ectopic calcification.”<-Fibroblast cells could be precursors to chondrogenic cells.

Here you can see the effects of teraparatide, it can increase height of the spine but also can cause scoliosis:


teriparatide effects

Height-Not all HGH and IGF-1

Short and tall stature: a new paradigm emerges.
Full Study->jeffreybaron study

“In the past, the growth hormone (GH)-insulin-like growth factor 1 (IGF-1) axis was often considered to be the main system that regulated childhood growth and, therefore, determined short stature and tall stature. However, findings have now revealed that the GH-IGF-1 axis is just one of many regulatory systems that control chondrogenesis in the growth plate, which is the biological process that drives height gain. Consequently, normal growth in children depends not only on GH and IGF-1 but also on multiple hormones, paracrine factors, extracellular matrix molecules and intracellular proteins that regulate the activity of growth plate chondrocytes. Mutations in the genes that encode many of these local proteins cause short stature or tall stature. Similarly, genome-wide association studies have revealed that the normal variation in height seems to be largely due to genes outside the GH-IGF-1 axis that affect growth at the growth plate through a wide variety of mechanisms. These findings point to a new conceptual framework for understanding short and tall stature that is centred not on two particular hormones but rather on the growth plate, which is the structure responsible for height gain.”

Regulation of growth plate function.Note extracellular fluid is listed as a factor and extracellular fluid flow can be modified by LSJL.

This page lists genes that cause mutations to linear growth.  This page lists other mutations and their effects on stature.

“the vast majority of children with short stature do not have a well-substantiated defect in the GH–IGF-1 axis.”

“Many new genes have been identified that, when mutated, result in short stature or tall stature, the majority of which do not participate in the GH–IGF-1 system”

“Estrogen has complex effects on the growth plate, not only altering growth rate, but also accelerating the loss of progenitor cells in the resting zone and thereby speeding up the developmental program of growth plate senescence, which causes early cessation of growth”

“tumour necrosis factor, IL-1β and IL-6 act directly on growth plate cartilage to suppress bone growth”

“fairly low doses of ionizing radiation, such as a single dose of 10 Gy, can impair longitudinal growth. Mechanical compression across the growth plate also impairs the elongation of bones, which is partly due to decreased enlargement of hypertrophic chondrocytes.”<-Chondrocyte hypertrophy size increases due to lsjl.

“FGFR-3 signalling negatively regulates growth by decreasing proliferation in the proliferative zone, decreasing production of the extracellular matrix, accelerating the onset of hypertrophic differentiation and decreasing the size of the hypertrophic chondrocytes”

“~2% of children who present with idiopathic short stature have mutations in NPR2, Conversely, overexpression of CNP or activating mutations in NPR2 result in tall stature.”

” Binding of CNP to NPR2 stimulates the guanylyl cyclase activity of the receptor, thereby increasing synthesis of cGMP, which activates the type II cGMP-dependent protein kinase”

“Of individuals presenting with idiopathic short stature, 2–15% have mutations in SHOX, with the exact percentage depending on the study. Conversely, increased copies of SHOX are associated with tall stature in individuals with Klinefelter syndrome and other types of sex chromosome aneuploidy”

“Sotos syndrome (characterized by tall stature) is associated with decreased activity of the Ras–MAPK pathway.”

“Ras, a small GTPase, signals via MAPK cascades to phosphorylate numerous cytoplasmic and nuclear proteins, regulating cell proliferation and differentiation.”

“heterozygous mutations in DNA methyltransferase 3A (DNMT3A) cause tall stature, a distinctive facial appearance and intellectual disability.”

“heterozygous mutations in EZH2, which encodes an enzyme that specifically methylates lysine residue 27 of histone 3 (H3K27, which is associated with transcriptional repression), are associated with Weaver syndrome (characterized by prenatal and postnatal overgrowth and a markedly advanced bone age).”

There’s a lot more covered in the study than I mentioned here.  The full study is worthwhile to read.