Monthly Archives: December 2013

BSP(Bone sialoprotein)

LSJL upregulates Bone sialoprotein.

Loss of bone sialoprotein leads to impaired endochondral bone development and mineralization.

“Bone sialoprotein (BSP) is an anionic phosphoprotein in the extracellular matrix of mineralized tissues, and a promoter of biomineralization and osteoblast development. Previous studies on the Bsp-deficient mouse (Bsp-/-) have demonstrated a significant bone and periodontal tissue phenotype in adulthood.  Early endochondral ossification in the Bsp-/- mouse was studied. Embryonic day 15.5 (E15.5) wild-type (WT) tibiae showed early stages of ossification that were absent in Bsp-/- mice. At E16.5, mineralization had commenced in the Bsp-/- mice, but staining for mineral was less intense and more dispersed compared with that in WT controls. Tibiae from Bsp-/- mice also demonstrated decreased mineralization and shortened length at postnatal day 0.5 (P0.5) compared to WT bones. There was no detectable difference in the number of tartrate-resistant acid phosphatase-positive foci at P0.5, although the P0.5 Bsp-/- tibiae had decreased Vegfα expression compared with WT tissue. Due to the shortened tibiae the growth plates were examined and determined to be of normal overall length. However, the length of the resting zone was increased in P0.5 Bsp-/- tibiae whereas that of the proliferative zone was decreased, with no change in the hypertrophic zone length of Bsp-/- mice{So BSP can possibly aid in the transition of resting zone chondrocytes to proliferative chondrocytes}. A reduction in cells positive for Ki-67, an S-phase cell-cycle marker, was noted in the proliferative zone. Decreased numbers of TUNEL-positive hypertrophic chondrocytes were also apparent in the Bsp-/- tibial growth plates, suggesting decreased apoptosis. Expression of the osteogenic markers Alp1, Col1a1, Sp7, Runx2, and Bglap was reduced in the endochondral bone of the neonatal Bsp-/- compared to WT tibiae. These results suggest that BSP is an important and multifaceted protein that regulates both chondrocyte proliferation and apoptosis as well as transition from cartilage to bone during development of endochondral bone.”

From this information though we can’t be clear whether the loss of BSP slows down growth or results in alteration of adult bone length.

“Overexpression of BSP downstream of a CMV promoter (CMV-BSP) in vivo resulted in mice that are significantly smaller than their wild-type controls. There are also defects in their endochondral bones, with aberrant growth plate formation and development. The growth plates of the CMV-BSP mice have no change in overall length, although there was an increase in hypertrophic zone size and number of hypertrophic chondrocytes. At 8 weeks of age, the mice also show a decrease in the proliferative zone length. This suggests that the overexpression of BSP promotes terminal differentiation in these chondrocytic cells.”<-So mice with overexpression of BSP had shorter bones.  Perhaps lower levels of BSP is beneficial as growth may be slower but take place over a longer period of time.

“these CMV-BSP mice have increased numbers of osteoclasts in their trabecular bone, suggesting that BSP promotes osteoclast formation”

“Mice deficient in BSP (Bsp−/−) also have decreased long bone length and cortical bone thickness relative to wild-type (WT) mice at 4 months of age. Conversely, they have a higher trabecular bone density than WT mice, but an apparent lower rate of bone turnover. This decrease in turnover could be due, in part, to a decrease in osteoclast numbers.”

“lack of BSP results in delayed patterns of bone development, which can be attributed, in part, to the reduction in the proliferation and apoptosis of the chondrocytes within the growth plate.”<-Delayed development not necessarily reduced development.

” The RP fraction was assayed for expression of developmental markers of chondrocyte development Alp1, Col2a1, Acan, Runx2, and Sox9 and no differences were seen between genotypes”

Review On The Grow Taller With Shin Lengthening Website

Review On The Grow Taller With Shin Lengthening Website

Shin LengtheningRecently one of the regular readers of the website expressed a desire for me to take a look at a website they have found entitled They did state that there seems to be nothing being sold on the website so it seems to be not doing any type of hard sell on anything.

I decided to spend about 30 minutes looking over the content and articles on the website. From checking over the sitemap part, it seems that the website has a total of 20 pages. I looked over the title of the pages hoping to find something that was new and not already stated in most other “grow taller” websites.

I chose not to care too much about the design and look of the website, since those things are not important.

Overall Conclusion

This website has always been designed to be based on scientific principles. We put extra effort in doing the research and making sure the science makes sense. We hoped that the Grow Taller With Shin Lengthening website would do the same. Overall, I felt that the website is lacking in serious content. From checking the Facebook Page that associated with the website, it seems that the website was created back in 2012.

The main claim made by this Dennis guy is that he managed to grow 7 inches taller in the years of what would be considered puberty for most adolescents. He claim he grew 3.5 inches in his shins alone which took him 3 years to do. He refer to a type of exercise known as Chaos Training, which I have on idea what that is. It took me a few minutes to realize that Chaos Training is similar to the P-90X workouts that are becoming really trendy in recent years. The exercise is intermittent but highly explosive. The idea is to do bursts of high aerobic and anaerobic exercise to get the testosterone increased. I would guess at this point he is just talking to the guys who are reading his website. The increase in testosterone would correlate to a similar spike in GH release.

We have shown in previous posts that males who have an above average level of testosterone level in their system before adolescence do seem to end up with a higher than average final adult height. However too much of the hormone would lead to stunted growth. It is a really tricky thing.

He also claims that if you stretch out your abdominal area you will be able to lengthen your torso by 3-5 inches as well just from stretching out the cartilage. 3-5 Inches??

Other things he suggest is to try dry swimming, the cobra stretch yoga position, and to go cycling. His suggestion that you should raise the height of the chair seat of the exercise cycle is something dozens of other “grow taller gurus” have suggested. There have been no results from this method, except maybe a few weak or injured knees.

The only section that was worth looking into was about how you can possibly lengthen the shins, which is just the front side of the tibia bones in the lower leg portion.

The main argument that is made by this Dennis Raney guy is that in Ancient China there was the practice of binding small girls feet to make them smaller. This shows that through excessive loading on the bone, the bone can be made to deform in the shape and angle one desires. The problem with this argument is that the Chinese Foot Binding is done to LITTLE GIRLS, NOT GROWN WOMAN. When you are young, the tarsals and meta-tarsal bones in the feet also have cartilage in them. The reason you can bind those feet to make them disfigured is that the binding is started when a girl is around 4-8 years old, much before her bones reach maturity, and the cartilage disappears. Try this practice of binding on a modern chinese woman’s feet and you would find that the feet can no longer bend to the will of just cloth. You might even get a slap over the face for being old fashioned and too traditional in thinking about the role of women.

It even states in the Wikipedia article that the binding of feet started even earlier, around the age of 2-5. (source)

In addition, I would like to quote how he defines the term puberty…

“Puberty is the stage in life when boys or girls become sexually mature. It is a stage of bodily change that usually happens between ages 10 and 14 for girls and ages 12 and 16 to 17 years for boys. Puberty affects boys and girls differently.”

Does this mean that this guy thinks that puberty is supposed to be over by the time we are around 17 years old?

I would guess due to just normal growth process and the variations of the distribution curve, for this person he probably kept growing past the age of 17. Growing in height at 19-21 is NOT unheard of. He went from 5′ 4″ to 5′ 11″. If I was to try to explain how he got so much taller, I would say he experienced a phenomena known as catch-up growth. Catch-Up Growth is what happens to the body of a adolescent who experienced slower growth rates earlier in their puberty years. The body goes into a sort of overcompensating process to increase the longitudinal growth of their bones to even out the growing line. For more understanding, we suggest reading the study Catch-up growth: definition, mechanisms, and models.

His claim on 7 inches over 3-4 years is not unreasonable. The Grow Taller Guru Lance Ward claimed that he grew about the same amount when he was around 20-21 as well. I would just put these guys growth process as an outlier, a data point in the overall set which is out of the ordinary. Growing taller for a male in their 20s is not normal but not impossible.

(Note: As a general rule I don’t link to websites which I don’t believe in. It is well known among people who work on the internet that putting in a link means that the website you link to gets more authority in the eyes of the search engines algorithms, especially what are known as high PR related websites).

Electromagnetic Stimuli Will Increase Ossification and Make Bones Longer, Big Breakthrough!

Electromagnetic Stimuli Will Increase Ossification and Make Bones Longer, Big Breakthrough!

Electromagnetic Stimuli Will Increase OssificationNote #1: I will be doing a podcast episode specifically about this post because it is so influential. There will be a lot of name dropping (sorry about that) and for all of you guys out there doing serious research just like me and Tyler, help out the cause and look into the links and studies which I will link below. I am just one person and research all of the leads. In some ways we ARE detective, trying to piece together puzzles to figure out what all of the data and study results actually mean. You can check out all of the older podcast episodes by CLICKING HERE!

Note #2: There is actually a section on Biological Immortality in this post. I will go into a little bit of detail on that as well to show that in my own quest to figure out how to grow taller, I might have also accidentally stumbled upon another interesting scientific endeavor, which is to figure out the mechanisms on how biological immortality is possible..

Articles like this don’t come every day, and this post I feel will be one of the most important posts I have ever written for the website, if not the biggest in terms of breakthroughs, because this is huge. The only other article that comes close might be the study where I showed that synthetic human epiphyseal cartilage had already been created more than a decade ago

Salamander Limb RegenerationRecently I finished reading the book The Body Electric by the late Dr. Robert O. Becker. (You can get the PDF for the book just by clicking the link above which is around 21 MB)  In the book Dr. Becker talked about how he would be able to stimulate lower level forms of organisms, specifically salamanders to regenerate entire limbs.

I also managed to upload for you guys to download the PDF for the book Electromagnetism and Life written by Dr. Becker and his assistant at the time Andrew Marino. Click the link above to download.

So the Beginning of this post Starts 3 Days Ago…

At the end of Dr. Becker’s seminal book, his tone turns angry as various government agencies and departments try to shut down his lab and his research. Something that is not well known is that the primary job of a professor is NOT to teach, BUT TO DO RESEARCH. To do research you need to have the money to buy all the lab equipment. That is why one of the main jobs of professors is to write grants to government agencies begging for money.

Robert Becker started to make a lot of enemies back in those days. He was actually interviewed on 60 Minutes along with his assistant at the time, an assistant/post-doc named Andrew Marino. They talked on the interview about the harmful effects of EMFs, which is short for Electromagnetic Fields.

It turns out that back in the 1980s and early 90s Becker was getting a lot of press from his research on the harmful effects of EMFs. I personally remember that there was even a movie starring Eddie Murphy called The Dinstinguished Gentlemen where he plays a conman-turned-politician who goes to a small town to see young kids who developed Cancer from living so close to high power lines. That was big news 20s years ago.

Andrew Marino LSUSo I finished the ending of the book, and decided to track down this Dr. Marino. It turns out that almost immediately after Dr. Becker’s lab was disbanded by the various government agencies he was fighting against back in the 80s and early 90s, Dr. Marino went to Louisiana State University, and has been there for at least 20 years now. Currently he holds the titles of ProfessorDirector of Research @ the Division of Sleep Medicine, Department of Neurology, Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA. Click Here to get to his faculty profile on the LSU website.

It seems that Dr. Marino actually writes his own blog and documents his personal thoughts. I found one of his older posts from back on Sept 9, 2011 (Click here to get to it) which was entitled “The 3 B’s and the Scientific Outlook“. The three B’s he is referring to are…

  1. Dr. Carl Brighton M.D, Ph. D. – I’ve talked extensively about him already. Still alive and only gave one email back in response, which was the first one. Never responded back to me ever again.
  2. Dr. Robert O. Becker M.D. – He is starting to become one of my scientific heros. He died fairly recently back in 2008. He was actually nominated for the Nobel Prize twice!
  3. Dr. C. Andrew L. Bassett M.D. – He was an orthopedic surgeon associated with Columbia University for a couple of decades. I tried to find him but found that he passed away back in 1994. Click here to read an article I found detailing his work and death

I clipped the cover of the the post for you guys to see. You can see what they looked like

Dr. Carl Brighton

So far I have only researched the work on Dr. Brighton and Dr. Becker. The reason why I have become almost obsessed with Dr. Becker’s work is because my 2nd Project, which I am focusing 90% of my energy on right now, is in doing research on the medical benefits of sending various types of electrical stimuli into the body.

Becker had evidence that it might be possible for humans to regenerate limbs from using DC current!

So I did two things…

I found the email to Dr. Marino on his website which is – I emailed Dr. Marino and asked whether he is willing to come on the website and let me interview him on the research he was so passionate about two decades ago. The fact that he writes about Dr. Becker even almost 20 years after the lab and research shows that he obviously still cares a lot about the work. He has currently not emailed me back. If he says yes, I will record out interview and place it as a podcast episode. Woohoo!

I once said that if I could get Dr. Brighton on the podcast for an interview it would be the greatest coupe for the website and our research. Since he never responded back after the 1st response which he did give, I guess it is never meant to be. Within the “3 Bs” he is the only one who is still alive!

When he passes away, he will take all of the research and work with him. Of course I am talking about the unpublished works, the lab experiments which didn’t work. Those are the one which are probably more worth studying. We know from business and academic research that most of our ideas and hypothesis don’t work out. In academics, you only publish the success stories. Your failures and failed lab tests are almost never recorded. That type of information can only be obtained through interview and questioning.

I sort of realize that Dr. Marino is still angry at Brighton since Brighton promoted EMFs but Becker was very much against EMFs. Brighton & Bassett said that EMFs were completely safe but Becker cautioned about it since his groups’ research showed otherwise.

There was a 2nd blog post published in Sept 19, 2011 entitled The 3 B’s and Electrical Stimulation of Bone Growth“. I also took the liberty of clipping the blog’s entry to show another picture of Bassett with Brighton.

Electrical Stimulation of Bone Growth

So since Mr. Marino has not responded back even after 3 days, there was only thing I could do.

I started to do research on the type of work that Bassett worked on. What I found was astounding!!

In the previous post, which was just my usual monthly update, I had said that there is a guy named Kronberg which had figured out to make a device for bone growth, in terms of bone density increases. That device will cause increased ossification but the medical benefit is reduced chances for bone fracture and reversal of the onset of osteoporosis from old age.

What Dr. Andrew Bassett did was much more in depth. Just from one source alone, it captures what I have been suspect of for a long time. Using PEMF stimulation does absolutely work in bone growth, and NOT JUST IN TERMS OF BONE DENSITY, but has an effect on cartilage tissue/cells as well!

From the 1994 New York Times article documenting his passing from a brain tumor, I quote what was written & summarized about Dr. Bassett’s 20 years of work…

“In 1978, after 20 years of investigation and a five-year study of outpatients, he reported that electromagnetic waves could be used to heal fractured limbs. He developed devices that put to work the discovery that cell functions within the organism could be controlled by external electromagnetic fields….”

I looked on PubMed, and Google Patents, and other archives to see what Bassett had published and invented back in the 1980s.

Finding #1: Bassett founded two companies, Electro-Biology Inc and Osteodyne Inc.

Electro-Biology Inc was eventually bought out by BioMet back in 1988. The current medical devices sold is the EBI’s OsteoGen™ Bone Growth Stimulator. This is from Bassett’s research. Another model is the SpinalPak spine fusion stimulators. I actually wrote a review on the device in an older post from last year.

This hows that all of the bone growth and fracture healing devices that BioMet currently sells are all derived from Bassett’s work from 20 years ago.

Finding #2: When I used google to find information on Osteodyne Inc, the results showed certain patents Bassett had published.

  • Method and device for treating bone disorders by applying preload and repetitive impacts – US 5484388 A
  • Device for treating bone disorders – EP 0427732 B1
  • Method and device for treating bone disorders – WO 1996003104 A1
  • Method and device for treating bone disorders characterized by low bone mass – US 5046484 A
  • Biomechanical-stimulation apparatus and method for bone regeneration – EP 2052708 A1
  • Non-invasive method and means for in-vivo bone-growth stimulation – WO 1993024092 A1

Of course those above are just his patents. The list below are his studies and papers published which I found on Pubmed.

All of these studies showed that if you applied DC Current in a frequency and intensity of just a reasonable level (often about 1-30 Amps), the bones can be stimulated for increased ossification. Some of the titles of the studies give it away immediately.

It also seems like Bassett & Becker collaborated on a few papers together. Their research was just sending electrical signals to the bone in a lab animal in vivo.

This G. Cochran guy is someone that I realized collaborated extensively with Bassett

What we have to realize is that the internet was not available back in the 80s or early 90s for most people. Only people who were associated with DARPAnet and the Department of Defense knew anything at all about how computers and connections worked back then.

If anyone would have been doing research to figure out height increase back then, they did not have the types of resources which we have now. There was no way that a lay person, or amateur medical researcher would be able to find this type of information. Now in the year 2013 we as amateur scientific researchers can use the power of the internet, the WayBack Machine, Google Scholars, and Patent Records to find all of this type of information which might have been recorded even 50-60 years ago.

Note: A lot of these studies are referenced and mentioned in the physician handbook Development And Growth edited by Geoffrey Bourne. I would love to get a copy of that book to go over the literature of what is written on bone fracture healing techniques and compare what I have found with that book.

So the obvious question for anyone who has done any type of research would be…

How does this information help us grow taller?

The obvious rebuttable is would be this – while E&M field generation could make bone fractures and non-unions heal, that DOES NOT mean that the bones get longer. Remember, every single device or pill (like using statin derivatives) lead to bone density or bone mass increase, NOT volumetric increase where the bone gets longer!!

Bones sort of have natural tendency to heal. Bones are actually one of the only tissue in the human body that has regenerative abilities. If you cut the heart, the wound will not heal over. A Bone fracture does. That is what makes it so special compared to the other tissues in the human body.

Based on The Body Electric book, I have learned that the locations where the bones actually start to heal is from two main places…

  1. The intermedullary cavity, which stores the bone marrow in any long bone in the human skeletal system.
  2. The periosteum – It seems that you always want to keep the periosteum intact.

The cortical bone and cancellous bone part DOES NOT heal, but those two areas.

What we really want to do is get the cartilage tissue in the bones to start to expand. To do that, the two chondrocyte processes of proliferation & hypertrophy must go together.

So What Can the Average Person Do?

This is a proposal which I think would be a very revolutionary idea. A person would need the following items.

1. A set of acupuncture needles that have been covered in silver metal.

2. Cooper wiring cord to connect the needle to the power source.

3. The power source

4. A way to change the frequency and the intensity of the electrical current.

The first thing that would be needed are silver/metal needles, similar in shape and size to what you see what acupuncturists use.

Robert Becker in the book talks about his research into the TCM practice of acupuncture and stated that his theory was that the needles when used on various pressure points in the body had some type of effect on

The other big thing is that from the research of Becker (and the Physicist Bob Beck, inventor of the Beck Protocol) it was found that silver has some extremely positive effects on bone growth. If you attach a silver sheet over a bone wound which has some serious infection and send negative potential (from a DC source) through the copper wiring it is connected to, the silver has some type of anti-microbial effect. Silver promotes bone growth and kills microbes. Of course it has been known for thousands of years since the ancient Chinese and Greeks that wound infections seem to heal faster from silver contact. Seriously Guys, Read the Book!!

I say that the acupuncture needles need to be covered in a silver layer because there will be some invasive techniques involved. Becker used electrodes that were completely silver, but I am not sure where a normal person would ever be able to buy 100% silver acupuncture needles.

Power Source – As for the Power Source, most people including Bassett, Brighton, and Becker all promoted DC Current although I am still in the camp believing in AC. Maybe even an ordinary AAA battery with a 1.5 Volt might be enough. What I do know is that while Brighton and his lab assistants used 10,000-20,000 NanoAmps per centimeter of silver electrode to get a response from the bone tissue, Becker and his lab assistances got the same results using just 100-200 NanoAmps (per centimeter of electrode) emitted. As for the volts – It seems that beyond just 1 Volt, Brighton’s research found that the DC power source seemed to have some type of damage to the tissue. That was too much. That means that we might not need a device that needs more than just 1 regular AAA battery. However, we do have to talk about the resistors.

Here was Dr. Becker’s reasoning on why acupuncture worked, which I paraphrased from his words in the book…

He thought the acupuncture meridians were electrical conductors that carried injury messages to the brain. Any current grows weaker with distance, due to resistant along the transmission cable line. This is basic Electrical Engineering knowledge.

The smaller the Amp or Volts, the faster the current dies out. That is why EEs have built current amplifiers along the cable line, to bring the current back up. Becker thought that those “chi” points aka acupressure points or “ki” points were the body’s natural amplifiers which boosted up the signal of current flowing down a nerve which is slowly lost due to natural resistance. So basically an acupuncture point was in analog a DC Power source or booster. Becker got a new assistant with Maria Reichmanis, and they showed that the traditional chinese medicine meridian lines were indeed conducting current, and the polarity of the readings showed  that for each acupuncture point was positive in polarity compared to the surrounding region. Each acupuncture point had their own electrical field being emitted. What was ultimately guessed by Becker was that the entire theory of acupuncture can be explained by the fact that the peri-neural cells were carrying current. That is what a metal needle stuck in would disrupt.

So the use of acupuncture needles will be three fold…

  1. They act as electrodes – we will connect them to a power source.
  2. They will be covered in silver, which is anti-microbial, and has bone generating abilities.
  3. They stimulate acupuncture points, which Becker seems to show are just peri-neural cells

What is a normal person supposed to do?

  1. Get the right type of silver electrode/ acupuncture needle.
  2. Find a normal conductive wire.
  3. Use a normal DC battery.
  4. Get an acupuncture chart – You want to get the dorsal region right above the SP9 point.

For proof that acupuncture works, read the PubMed study “Trigger point acupuncture for treatment of knee osteoarthritis“. They had used trigger point acupuncture therapy.

I personally would guess that what a person can start doing is first they stick the needle/electrode to just the area right above the epiphyseal plates. We found from a few old studies that if you cut a hole in the diaphysis of long bone and use an awl to cut apart the blood vessels, the longitudinal bone growth gets increased slightly. This is due to the fact that long bones have separate groups of blood vessels going to separate regions. There is a group of arteries going only to the epiphysis and another group supplying the blood to the diaphysis. What I am saying is that we want to get the electrodes to be close to the blood vessels supplying nutrients to the epiphysis only. Why that is the case we are not sure yet.

What about the Frequency and Intensity that we need to adjust for?

This part will need to be edited for later. There are a lot of values for frequency, duration of session, number of sessions per week, and intensity of the current, flying around in the various studies. I’ll edit this section when I organize everything.

I am extremely confident that this technique will result in increased longitudinal growth for the long bones for people will intact epiphyseal growth plates. For children who wish to be taller, this would be the most cost effective alternative that is possible. 

Recently the company Biomarin has developed a drug known as BMN-111 which is supposed to treat dwarfism for children only (aka people still with growth plates) . You can read the article on the drug by clicking here. The way the drug works is through the stimulation of the CNP and through inhibition of the FGFR3 gene. We have already done research and understand the process of how CNP is stimulated through mTOR and Rapamyxin reasonably well. That treatment would cost over $50,000 for any parent who does not wish their kids to end up less than 5 feet tall. This I believe is a very reasonable alternative to Biomarin’s Dwarfism treatment, which would involve a lot of money. 

Tyler has been promoting both the LIPUS and PEMF technology for years, but I personally think he has put more emphasize on the LIPUS method. From this post, I am trying to give conclusive proof that it would be much better to be focusing and promoting the PEMF path.

Further Validation of This Concept:

I refer to the Patent – Methods for modulating chondrocyte proliferation using pulsing electric fields – US 20080039901 A1

This patent was published back in 2008 by James Kronberg, who I have mentioned before. You can find the same patent from Google Patents. He used a pulse burst mode and continuous mode for the signal shape and duration. From the abstract he states that you can increase cartilage repair from stimulating NO production….

“…Cartilage, bone and other connective tissue growth is stimulated in part by nitric oxide release through electrical stimulation and may be modulated through co-administration of NO donors and NO synthase inhibitors. The methods and devices described are useful in promoting repair of bone fractures, cartilage and connective tissue repair as well as for engineering tissue for transplantation”

The other articles are…

The abstract of this paper almost states in plain english that using a PEMF of just low intensity would be enough to stimulate the process of endochondral ossification. You can get an increase in cartilage molecules, glycoaminoglycan content , and extracellular matrix production.

Million Dollar Question – Will It help a person with closed growth plate grow longer bones?

I suspect at this point that for people with closed growth plates, there is a slight probability, with maybe a 5-10% chance of it having some effect.

This is due to three related studies I did find.

  1. Effects of pulsed electromagnetic fields on human articular chondrocyte proliferation.
  2. Millicurrent stimulation of human articular chondrocytes cultivated in a collagen type-I gel and of human osteochondral explants.
  3. Effects of pulsed electromagnetic fields on articular hyaline cartilage: review of experimental and clinical studies

I showed in a recent post that there are still chondrocytes in the adult human articular cartilage in the ends of the epiphysis. The post was Articular Cartilage At The End Of Epiphysis Do Growth Thicker Making Bones Longer (Big Breakthrough)”

This method would have some type of dramatic effect on the articular cartilage as well.

If we remember that way that the tissues layers are aligned next to each other underneath the cartilage layer, we realize that there is a subchondral layer. That layer can be thickened using the correct implantation location of the silver electrode to stimulate both an increase in the cartilage ECM and the increased deposition of the bone layer.

This part still needs to be edited further.


The section on biological immortality

The finding was actually completely unexpected. In recent years, starting around the 2010-2011 time frame, there was  a lot of articles published about this biologically immortal living organism called the Hydra. We suggest the study/paper MORTALITY PATTERNS SUGGEST LACK OF SENESCENCE IN HYDRA or “Longevity Gene Makes Hydra Immortal and Humans Grow Older

It would turn out that when I was reading up on Becker’s book that the Hydra was mentioned very early on. I took the chance to clip a picture of the Hydra which was from page 34 of the book.

Biologically Immortal Hydra

A lot of the journalists who tried to cover the story and science on the Hydra seems to state that there is almost no research and information on the Hydra. Well they are wrong!

Becker and Abraham Trembley who lived 3 centuries were already studying this immortal creature. It seems that not only is the hydra biologically immortal, it might be completely immortal. Cutting the creature won’t kill it.

So not only can it live forever, it is almost nearly impossible to kill the creature even if you wanted to!!

It’s only weakness seems to be in the stalk. If the hydra cuts a leg or head off without the stalk, then it won’t regenerate. however if the head or leg cut off has any part of the stalk, the cut pieces will all eventually regenerate back into a whole intact body.

Hydra Regeneration

This is where the scientists right now who are studying why senescence seems to go in reverse for Hydra have not made the connection yet. There is even more to the Hydra that they realize.

What are the implications?

My research for this website was always towards height increase, the manipulating of tissue engineering, advanced electromagnetic stimuli, and such for a cosmetic gain. This finding I saw from reading over books written by researchers who have been criticized.

There has been a lot of talk in the medical community about the discovery of the FoxO Gene and that has sort of gotten people excited. The way that the Hydra reproduces is through budding. For further reading on the Hydra and immortality, read the article Could this immortal hydra polyp inspire advanced rejuvenation therapies for humans?

Slc26a2 height increase gene

Slc26a2 is involved in sulfur intake by chondrocytes and mutations in Slc26a2 can cause Diastrophic Dysplasia, which causes chondrocyte undersulfation and reduces height.  Read more about DTD here.  A supplement that upregulates Slc26a2 expression could be an alternative to increasing IGF-1 expression and given that DTD is a disease treatments will be developed that may be applicable to people with operating normal growth plates to grow taller.

Multiple Roles of the SO42-/Cl-/OH- exchanger Slc26a2 in Chondrocyte functions.

“Mutations in the SO42-/Cl-/OH- exchanger Slc26a2 cause the disease diastrophic dysplasia (DTD), resulting in aberrant bone development and therefore skeletal deformities. DTD is commonly attributed to a lack of chondrocyte SO42- uptake and proteoglycan sulfation. However, the skeletal phenotype of patients with DTD is typified by reduction in cartilage and osteoporosis of the long bones. Chondrocytes of patients with DTD are irregular in size and have a reduced capacity for proliferation and terminal differentiation. This raises the possibility of additional roles for Slc26a2 in chondrocyte function. Here, we examined the roles of Slc26a2 in chondrocyte biology using two distinct systems: mouse progenitor mesenchymal cells differentiated to chondrocytes and freshly isolated mouse articular chondrocytes differentiated into hypertrophic chondrocytes. Slc26a2 expression was manipulated acutely by delivery of Slc26a2 or shSlc26a2 with lentiviral vectors. Slc26a2 is essential for chondrocyte proliferation and differentiation, and for proteoglycan synthesis. Slc26a2 also regulates the terminal stage of chondrocyte cell size expansion{thus perhaps extra Slc26a2 could increase height}.”

“Slc26a2 functions as a SO42-/Cl-/OH exchanger that is exquisitely regulated by extracellular Cl-”

“Slc26a2 expression is highly enriched in the long bone proliferating zone.”  Slc26a2 is minimally expressed in the resting zone but moderately expressed in the hypertrophic zone.  Thus, perhaps Slc26a2 plays a role in chondrocyte differentiation.

“Proteoglycan sulfation depends on SO42- uptake by chondrocytes. The SO42- transporter Slc26a2 mediates most SO42- uptake by chondrocytes”

IGF-1 activates Slc26a2.

“Knockdown of Slc26a2 strongly inhibited the effect of IGF-1 on proteoglycan synthesis, and IGF-1 had a very modest effect on proteoglycan synthesis in cells overexpressing Slc26a2.  Inhibition of PI3K by LY294002 inhibited the effects of both Slc26a2 and IGF-1.”<-Thus Slc26a2 is highly responsible for the rate of proteoglycan synthesis.  High IGF-1 can make up for a Slc26a2 deficit and vice versa.

“treatment with IGF-1 and overexpression of Slc26a2 increased proteoglycan synthesis, whereas knockdown of Slc26a2 inhibited synthesis.”

“IGF-1 and Slc26a2 increased chondrocyte size, with a combined effect smaller than the additive effect. PI3K activity was necessary for both IGF-1 mediated and Slc26a2 mediated chondrocyte volume expansion.”

“the function of Slc26a2 in chondrocytes cannot be compensated for by any of the other SLC26 SO42- transporters”

“Slc26a2 is constitutively phosphorylated by PI3K”

Chondrogenic Induction with only Mechanical Stimulation

The following study provides a new paradigm in inducing longitudinal bone growth in adults.  Cartilage versus Bone cell differentiation is movement based and bone is stiff.  If bone tissue was exposed to more deformation(like that which occurs at a joint) then cartilage differentiation could occur and neo growth plate formation is one possible result.  LSJL is one possible mechanism for inducing this deformation as bone is more malleable in the lateral direction than the axial direction that occurs more frequently in physiological loading.

This paradigm opens the possibility for other ways to deform the bone to encourage chondrogenic differentiation within the bone to form new growth plates.

Pressureless mechanical induction of stem cell differentiation is dose and frequency dependent.

“Pressureless, soft mechanical stimulation precipitated by the cyclic deformation of soft, magnetic hydrogel scaffolds with an external magnetic field, can induce chondrogenesis in mesenchymal stem cells without any additional chondrogenesis transcription factors (TGF-β1 and dexamethasone). A systematic study on the role of movement frequency revealed a classical dose-response relationship for human mesenchymal stem cells differentiation towards cartilage using mere mechanical stimulation.”

“Diffusion, local release of growth factors and concentration gradients shape an organism’s 3D structure. Unfortunately, liquid filled pockets and particularly movement destroy such local information pattern (e.g. in a joint)

cartilage interface

“Hip joint (anatomy, a) with a local concentration profile of a differentiation factor secreted at the interface of the cartilage if the joint is not moved (hypothetical, b) and under physiologic movement (c). Diffusion alone usually results in rather steep concentration gradients (static situation). Movement induces liquid mixing through convection (synovial fluid) flattening concentration profiles. ”

” Chondrocyte growth has been associated with demanding physical input such as cell deformation, hydrostatic pressure gradients, fluid flow, streaming currents and physicochemical changes”<-LSJL could potentially alter all of these.

“Without adequate traumatic surgery or fixation, pseudoarthrosis may occur at non-fixed bone fracture sites. Continued movement of such non-fixed primary callus then promotes local stem cell differentiation into cartilage and ligament tissue instead of bone tissue.”<-Mimicking this concept without the fracture may enable neo-growth plate formation.

“If a joint is not moved anymore (e.g. due to arthritis), even an otherwise fully functional joint stiffens (ankylosis) and the tissue locally transforms into a bone-type material”

So movement tends to encourage cartilage growth whereas lack of movement encourages bone growth.  The bone changes in length constantly as noted by the definition of microstrain(which measures strain as change in bone length).  So natural movement is not enough.  However, something like LSJL may mimic such movement via lateral bone deformation as bone is more malleable from a lateral direction than an axial direction.

Bone bending and twisting does occur in a natural environment.  It’s just that the natural loads that occur are not enough movement to encourage cartilage over bone stem cell differentiation.

“For the impact of mechanical stimulation at different intensities regarding to differentiation, the stimulus was performed in 30 min cycles every 1.5 h for 8 h per day during 3 weeks (daytime activity, total number of cycles = 12600, 2016 and 672 respectively) for each group (n = 3). Cell culture samples were taken after 1, 2 and 3 weeks, respectively.”

“The only difference between differentiating and non-differentiating hMSC seeded on the scaffolds used here, was the application of the external magnetic field and the resulting reversible, soft deformation of the scaffold. “

The Connection Between Relaxin And Possible Height Increase

Me: Ever since we started to find stories and posts on the internet mothering and pregnancy boards where women have come forward to state that their height increase from their pregnancy, the topic of relaxin as a possible height increase hormone has been raised. Tyler has recently done a post on the subject (link HERE).

Tyler: Added a new study directly below.  The most likely cause of the pregnancy height gain some females experience is due to increase in tendon length as there doesn’t seem to be any logical connection between relaxin and longitudinal bone growth.  Although the cattle pelvic height growth study seems to indicate that relaxin could somehow induce articular cartilage endochondral ossification results in pelvic height growth.

The effect of relaxin on the musculoskeletal system.

“Relaxin circulates during pregnancy emanating from the corpus luteum and placenta”

“In rodents, circulating relaxin peak concentrations at the end of pregnancy reach 100 ng/mL, two times greater than in human”

“seven known relaxin family peptides (RXFP) are structurally related to insulin which include relaxin (RLN)1, RLN2, RLN3, and insulin-like peptide (INSL)3, INSL4, INSL5, and INSL6″

” Relaxin alters cartilage and tendon stiffness by activating collagenase”

The study notes that Relaxin increases length in cartilage and tendons(Fig 2).

Relaxin has a role in osteoclastgenesis.

“Relaxin treatment in pregnant cattle increased pelvic width and height, but not in other joints such as wrist and knee”

“Relaxin appears to decrease knee articular cartilage stiffness through induction of collagenase-1, MMP-1, and MMP-3, which reduces collagen content and expression in fibrocartilaginous cells. Modulation of MMPs to loss of collagen by hormones may contribute selectively to degeneration of specific joints fibrocartilaginous explants facilitated by proteinases. The degradation of extracellular matrix in fibrocartilage is synergized by β-estradiol. Relaxin exerts its effect through binding to RXFP1 and RXFP2 receptors. The ratio of RXFP2 in knee meniscus of pregnant rabbits was shown to be more than RXFP1, which may indicate differential role of these receptors in the remodeling of fibrocartilage. Comparison of collagen content in articular cartilage of nonpregnant and pregnant rabbits showed that the total RNA levels and chondrocyte metabolism decreased during pregnancy.”

One study has noted that porcine relaxin is capable of modifying type II collagen expression in chondrocyte cells.  Another study found that relaxin increased tendon length so tendon length could be related to the height grain in pregnancy.

This is my attempt in trying to see whether any PubMd studies I have found would be worth mentioning in relevance to our research. I would cite and copy below 5 studies.

Analysis & Interpretation:

From the 1st study…

I would assume Tyler in his post used the study “Pelvic development as affected by relaxin in three genetically selected frame sizes of beef heifers”. The really interesting thing is that the study seems to show that the entire pelvic bone area increased in size. The term “primiparous beed heifers” refer to female cows which are going through their first birthing experience. The issue with the study is of course that cows were used than humans. Also it doesn’t really go and try to explain how it was possible that relaxin caused the increase in pelvic height increase or pelvic width increase. I am now cow expert so I don’t know whether the cows were still growing by the standards of humans. What I do know is that cows which are going through their first birthing experience can easily be young enough to still have growth plate cartilage so what we see in pelvic bone height can be just the excess release of whatever hormones female cows will go through when they are pregnant. From the controlled study we see that relaxin being introduced does cause clear increases in pelvic bone height and width.

For humans, we have to remember the way human female reproductive systems work. The indication that human females can grow is from the start of the menses cycle. The first menses is known as the menarche. Most girls would have their menarche during the puberty stage, but before the growth plates ossify completely. This means that before girls stop growing taller, they would develop the ability to have children and go through the human gestation process. In bovine, we would guess it is the same thing. In conclusion, from just this one study we can see that in cows, the hormone relaxin has the ability to increase pelvic height. Since the pelvic bone outline/structure does indeed contribute to the overall human height, the increase in pelvic height we see in young female bovine may be able to be translated to young human females. However the age values we find from the internet discussion boards and forum suggest that the relaxin is doing something more exordinary since the women are already even in their late 20s or 30s when they notice the height increase. If relaxin is just only a muscle relaxant, how does it actually make pelvic bone taller? Just something to wonder about right now.

{Tyler: “Pelvic height was determined by measuring the linear distance from the approximate midpoint of the dorsal surface of the symphysis pubis to the ventral surface of the prominent junction of the third and fourth segments of the sacral vertebrae”<-Doesn’t seem to factor in tendon length.

“pelvic growth during the last 10 days before parturition[child birth] can be modified by the intracervical administration of relaxin.”

“The mechanism of pelvic canal expansion in cattle is unknown. The pelvic canal may increase in area as a result of relaxation of sacroiliac ligaments, formation of interpubic ligaments, and modification of the pubic symphysis by transforming symphyseal cartilage[a type of cartilage joint] and bone{if it does this then relaxin could have very strong height increase potential}“<-growing from the pubic symphysis would be like growing from the articular cartilage which would have very nice height increase implications.  The study that discsusses this is called Dystocia in Cattle by LE Rice unfortunately I could not find this study.

From the 2nd study…

If we however look at the 2nd study I have linked, we can see that Relaxin, specifically the Relaxin Family Peptide Receptor 1 (RXFP1) can be the precursor to a mechanism which triggers many of the other pathways and hormones we have looked at before. including NO and cGMP. Somehow it disrupts the TGF-β1/Smad2 axis through a signal process that involves the ERK/pERK/NO/cGMP pahway. One set of proteins I have been trying to get to researching more into are the MMPs. Somehow the RXFP1 increase matrix metalloproteinase (MMP) expression. My knowledge on MMPs are nonexistent at this point so I can’t really breakdown what the abstract is really talking about. However the researchers would conclude with…

“These findings demonstrated that H2 relaxin signals through a RXFP1-pERK-nNOS-NO-cGMP-dependent pathway to mediate its anti-fibrotic actions, and additionally signals through iNOS to up-regulate MMPs; the latter being suppressed by TGF-β1 in myofibroblasts, but released upon H2 relaxin-induced inhibition of the TGF-β1/Smad2 axis”

So I guess the two main things to take away from this study is that the H2 relaxin seems to inhibit/suppress the differentiation of myofibroblast and also increase the gene expression of the MMPs.

From the 3rd study…

Relaxin seems to regulate the expression of two types of MMPs, MMP-9 and MMP-13. Relaxin is a type of ligand that attaches to a certain type of substrate or receptor which will accept it. There is two types of relaxin receptors described in this study, Relaxin family peptide receptors 1 & 2, called RXFP1 & RXFP2 respectively. It would seem that the RXFP1 is the one that has the real regulating power on MMP9 & MMP-13 expression while the RXFP2 doesn’t seem to regulate the MMP expression. There are quite a few pathways involved in the relaxin’s regulating ability including the PI3K, AKT, ERK, and a few other pathways or compounds which I am not familiar with at this time. The main thing to take away from this study is that relaxin has some regulating ability over the extracellular matrix and MMP expression.

From the 4th study…

From the 3rd study we learn that there seems to be at least two main relaxin receptors, called the relaxin family peptide receptors, RXFP1 and RXFP2. It seems that while the first receptor RXFP1 is for the actual relaxin compound in a ligand-substrate match, the 2nd receptor is for matching with something called insulin like peptide (INSL)3. Both of the receptors and thus both of the compounds increase the cAMP level but through two different pathways, also both going through a compound called a G-Alpha. At this point I don’t understand or know what most of the abstract is talking about but the key to understand is that the relaxin and another compound very similar to it both increases cAMP expression. Like what we find in the 2nd and 3rd studies, relaxin can increase the up-regulation of a few key compounds which we have looked at before in our research.

From the 5th study…

It would seem that this compound we have been looking at through the last 5 studies is well known as a an anti-fibrotic element. It is a peptide hormone that inhibits fibrosis of different types, but for this study specifically the cardiac fibrosis type. When you take TGF-Beta or Angiotensin Type II (Ang II) it would cause accelerated fibroblast differentiation into myofibroblasts. Relaxin seems to inhibit the differentiation of fibroblasts which are treated with Ang II, IGF-1, or TGF-beta. This was found from detecting that the expression of alpha-smooth muscle actin and collagen decreased. MMP-2 expression was also noted to have increased from relaxin under the presence of the Angiotensin II and TGF-beta.

The researchers would conclude with…

“These coherent findings indicate that relaxin regulates fibroblast proliferation, differentiation, and collagen deposition and may have therapeutic potential in diseased states characterized by cardiac fibrosis”


At this point, I would say that relaxin is something that I would need to study further since it and it’s receptors the RXFP1 and RXFP2 have some regulating function towards many types of the MMPs, MMP2, 9, and 13. It also have control over the expression of cAMP and fibroblast differentiation ability.

Biol Reprod. 1986 Mar;34(2):363-9.

Pelvic development as affected by relaxin in three genetically selected frame sizes of beef heifers.

Musah AI, Schwabe C, Willham RL, Anderson LL.


Purified porcine relaxin was administered into the cervical os on Day 278 of gestation to determine its effects on pelvic development in three genetically selected frame sizes of primiparous beef heifers. Heifers were categorized as small, medium and large frame based upon their genetic composition. Pelvic height, pelvic width and cervical dilatation were determined from Day 270 to 2 days postpartum. On Day 270, heifers were assigned at random to one of three treatments: vehicle control, n = 16; relaxin once (3,000 U), n = 14; and relaxin twice (2 times 3,000 U 12 h apart), n = 17. Each heifer-frame size was represented in each treatment. Relaxin caused marked increases in pelvic height and width, as well as in the rate of linear increase (cm/day) of these parameters (p less than 0.05). These linear increases in pelvic height were 510, 264 and 204%, and pelvic width, were 280, 213 and 204% of the respective pretreatment rates for small, medium and large heifers. The rate of linear increase in pelvic width was greater than pelvic height in all heifers, but maximal in small-frame heifers; relaxin attenuated these intrinsic differences. For small heifers, the rate of linear increase in pelvic width was 121 and 145% of increases for medium and large heifers, respectively, before treatment, and 160 and 200% after treatment. The rate of postpartum involution of pelvic width was -0.03, -0.36 and -0.50 cm/day and, for pelvic height, -0.02, -0.27 and -0.29 cm/day in small, medium and large heifers, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

PMID: 3955148     [PubMed – indexed for MEDLINE]    Free full text

From PubMed study 2 link HERE

PLoS One. 2012;7(8):e42714. doi: 10.1371/journal.pone.0042714. Epub 2012 Aug 22.
Relaxin signals through a RXFP1-pERK-nNOS-NO-cGMP-dependent pathway to up-regulate matrix metalloproteinases: the additional involvement of iNOS.
Chow BS, Chew EG, Zhao C, Bathgate RA, Hewitson TD, Samuel CS.

Florey Neuroscience Institutes, University of Melbourne, Parkville, Victoria, Australia.


The hormone, relaxin, inhibits aberrant myofibroblast differentiation and collagen deposition by disrupting the TGF-β1/Smad2 axis, via its cognate receptor, Relaxin Family Peptide Receptor 1 (RXFP1), extracellular signal-regulated kinase (ERK)1/2 phosphorylation (pERK) and a neuronal nitric oxide (NO) synthase (nNOS)-NO-cyclic guanosine monophosphate (cGMP)-dependent pathway. However, the signalling pathways involved in its additional ability to increase matrix metalloproteinase (MMP) expression and activity remain unknown. This study investigated the extent to which the NO pathway was involved in human gene-2 (H2) relaxin’s ability to positively regulate MMP-1 and its rodent orthologue, MMP-13, MMP-2 and MMP-9 (the main collagen-degrading MMPs) in TGF-β1-stimulated human dermal fibroblasts and primary renal myofibroblasts isolated from injured rats; by gelatin zymography (media) and Western blotting (cell layer). H2 relaxin (10-100 ng/ml) significantly increased MMP-1 (by ~50%), MMP-2 (by ~80%) and MMP-9 (by ~80%) in TGF-β1-stimulated human dermal fibroblasts; and MMP-13 (by ~90%), MMP-2 (by ~130%) and MMP-9 (by ~115%) in rat renal myofibroblasts (all p<0.01 vs untreated cells) over 72 hours. The relaxin-induced up-regulation of these MMPs, however, was significantly blocked by a non-selective NOS inhibitor (L-nitroarginine methyl ester (hydrochloride); L-NAME; 75-100 µM), and specific inhibitors to nNOS (N-propyl-L-arginine; NPLA; 0.2-2 µM), iNOS (1400W; 0.5-1 µM) and guanylyl cyclase (ODQ; 5 µM) (all p<0.05 vs H2 relaxin alone), but not eNOS (L-N-(1-iminoethyl)ornithine dihydrochloride; L-NIO; 0.5-5 µM). However, neither of these inhibitors affected basal MMP expression at the concentrations used. Furthermore, of the NOS isoforms expressed in renal myofibroblasts (nNOS and iNOS), H2 relaxin only stimulated nNOS expression, which in turn, was blocked by the ERK1/2 inhibitor (PD98059; 1 µM). These findings demonstrated that H2 relaxin signals through a RXFP1-pERK-nNOS-NO-cGMP-dependent pathway to mediate its anti-fibrotic actions, and additionally signals through iNOS to up-regulate MMPs; the latter being suppressed by TGF-β1 in myofibroblasts, but released upon H2 relaxin-induced inhibition of the TGF-β1/Smad2 axis.

PMID: 22936987 [PubMed – in process] PMCID: PMC3425563

From PubMed study 3 link HERE

Mol Cell Endocrinol. 2012 Nov 5;363(1-2):46-61. doi: 10.1016/j.mce.2012.07.006. Epub 2012 Jul 24.
Relaxin induces matrix-metalloproteinases-9 and -13 via RXFP1: Induction of MMP-9 involves the PI3K, ERK, Akt and PKC-ζ pathways.
Ahmad N, Wang W, Nair R, Kapila S.

The University of Michigan, 1011 North University Avenue, Ann Arbor, MI 48109-1078, USA.


We determined the precise role of relaxin family peptide (RXFP) receptors-1 and -2 in the regulation of MMP-9 and -13 by relaxin, and delineated the signaling cascade that contributes to relaxin’s modulation of MMP-9 in fibrocartilaginous cells. Relaxin treatment of cells in which RXFP1 was silenced resulted in diminished induction of MMP-9 and -13 by relaxin, whereas overexpression of RXFP1 potentiated the relaxin-induced expression of these proteinases. Suppression or overexpression of RXFP2 resulted in no changes in the relaxin-induced MMP-9 and -13. Studies using chemical inhibitors and siRNAs to signaling molecules showed that PI3K, Akt, ERK and PKC-ζ and the transcription factors Elk-1, c-fos and, to a lesser extent, NF-κB are involved in relaxin’s induction of MMP-9. Our findings provide the first characterization of signaling cascade involved in the regulation of any MMP by relaxin and offer mechanistic insights on how relaxin likely mediates extracellular matrix turnover.

Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

PMID:  22835547     [PubMed – in process] 

From PubMed study 4 link HERE

Mol Pharmacol. 2006 Jul;70(1):214-26. Epub 2006 Mar 28.
Relaxin family peptide receptors RXFP1 and RXFP2 modulate cAMP signaling by distinct mechanisms.
Halls ML, Bathgate RA, Summers RJ.

Department of Pharmacology, P.O. Box 13E, Monash University, Clayton, VIC 3800, Australia.


Two orphan leucine-rich repeat-containing G protein-coupled receptors were recently identified as targets for the relaxin family peptides relaxin and insulin-like peptide (INSL) 3. Human gene 2 relaxin is the cognate ligand for relaxin family peptide receptor (RXFP) 1, whereas INSL3 is the ligand for RXFP2. Constitutively active mutants of both receptors when expressed in human embryonic kidney (HEK) 293T cells signal through Galphas to increase cAMP. However, recent studies using cells that endogenously express the receptors revealed greater complexity: cAMP accumulation after activation of RXFP1 involves a time-dependent biphasic pathway with a delayed phase involving phosphoinositide 3-kinase (PI3K) and protein kinase C (PKC) zeta, whereas the RXFP2 response involves inhibition of adenylate cyclase via pertussis toxin-sensitive G proteins. The aim of this study was to compare and contrast the cAMP signaling pathways used by these two related receptors. In HEK293T cells stably transfected with RXFP1, preliminary studies confirmed the biphasic cAMP response, with an initial Galphas component and a delayed response involving PI3K and PKCzeta. This delayed pathway was dependent upon G-betagamma subunits derived from Galphai3. An additional inhibitory pathway involving GalphaoB affecting cAMP accumulation was also identified. In HEK293T cells stably transfected with RXFP2, the cAMP response involved Galphas and was modulated by inhibition mediated by GalphaoB and release of inhibitory G-betagamma subunits. Thus, initially both RXFP1 and RXFP2 couple to Galphas and an inhibitory GalphaoB pathway. Differences in cAMP accumulation stem from the ability of RXFP1 to recruit coupling to Galphai3, release G-betagamma subunits and thus activate a delayed PI3K-PKCzeta pathway to further increase cAMP accumulation.

PMID:  16569707   [PubMed – indexed for MEDLINE]      Free full text

From PubMed study 5 link HERE

Endocrinology. 2004 Sep;145(9):4125-33. Epub 2004 May 20.
Relaxin modulates cardiac fibroblast proliferation, differentiation, and collagen production and reverses cardiac fibrosis in vivo.
Samuel CS, Unemori EN, Mookerjee I, Bathgate RA, Layfield SL, Mak J, Tregear GW, Du XJ.

Howard Florey Institute, Gate 11, University of Melbourne, Parkville, Victoria 3010, Australia. c.samuel@hfi.unimelb.


Cardiac fibrosis is a key component of heart disease and involves the proliferation and differentiation of matrix-producing fibroblasts. The effects of an antifibrotic peptide hormone, relaxin, in inhibiting this process were investigated. We used rat atrial and ventricular fibroblasts, which respond to profibrotic stimuli and express the relaxin receptor (LGR7), in addition to two in vivo models of cardiac fibrosis. Cardiac fibroblasts, when plated at low density or stimulated with TGF-beta or angiotensin II (Ang II), accelerated fibroblast differentiation into myofibroblasts, as demonstrated by significantly increased alpha-smooth muscle actin expression, collagen synthesis, and collagen deposition (by up to 95% with TGF-beta and 40% with Ang II; all P < 0.05). Fibroblast proliferation was significantly increased by 10(-8) m and 10(-7) m Ang II (63-75%; P < 0.01) or 0.1-1 microg/ml IGF-I (27-40%; P < 0.05). Relaxin alone had no marked effect on these parameters, but it significantly inhibited Ang II- and IGF-I-mediated fibroblast proliferation (by 15-50%) and Ang II- and TGF-beta-mediated fibroblast differentiation, as detected by decreased expression of alpha-smooth muscle actin (by 65-88%) and collagen (by 60-80%). Relaxin also increased matrix metalloproteinase-2 expression in the presence of TGF-beta (P < 0.01) and Ang II (P < 0.05). Furthermore, relaxin decreased collagen overexpression when administered to two models of established fibrotic cardiomyopathy, one due to relaxin deficiency (by 40%; P < 0.05) and the other to cardiac-restricted overexpression of beta2-adrenergic receptors (by 58%; P < 0.01). These coherent findings indicate that relaxin regulates fibroblast proliferation, differentiation, and collagen deposition and may have therapeutic potential in diseased states characterized by cardiac fibrosis.

PMID: 15155573    [PubMed – indexed for MEDLINE]       Free full text