An Interview With Dr. Tarek El-Bialy On Using Ultrasound To Regenerate Tooth Dentin

A few days ago we got a comment by someone calling themselves Pete who linked to an interview given where the Canadian Dr. Tarek El-Bialy was being interviewed on his Ultrasound Device & technology to use to regrow teeth, but stimulating the root of the teeth.

Originally we had written about the technology in a very old post “Teeth Regrowth Using Low Intensity Pulsed Ultrasound, LIPUS“. The researcher we mentioned in that article was a Dr. Jie Chen, also in Canada who claimed that they could use Ultrasound to not just make bones heal fractures but make people grow taller, which was something that was edited into the Wikipedia article on LIPUS (Low Intensity Pulsed Ultrasound). Tyler really got into the idea of using LIPUS and really deep into LIPUS but I had written in the past year a few times showing multiple articles which disprove the idea that Ultrasound would ever lead to increased longitudinal growth in the long bones. Yes, LIPUS machines are already on sale at rather cheap prices, so if there was real evidence in articles and the scientific literature that shows the really small LIPUS devices does any even the slightest chance of increasing longitudinal growth, I would recommend the portable LIPUS devices on this website, since they are just around $30-$50 each, and may be even free if one has the right type of medical insurance.

In fact, I recently downloaded the PDF of an old article I had eluded to to reread the implications on what would happen if you continuously emit ultrasound in lab test rabbits growth plates. (Effect of continuous therapeutic ultrasound in rabbit growth plates).

However, the technology of LIPUS on nerve tissue and teeth regeneration is much more promising. As Dr. El-Bialy (Associate Professor of Orthodontics and Biomedical Engineering at the University of Alberta, Canada) seems to suggest, LIPUS can regenerate damaged nerves and root pulp.

You can download the MP3 of that interview wth Dr. El-Bialy Available Here

It may not be our primary focus, but learning about the science of teeth and regeneration of teeth might lead to less money we have to spend at the dentist office in the future.

This study doesn’t relate directly to height growth but it is by the scientists whose research was the foundation for LSJL.

In vitro and in silico analysis of an inhibitory mechanism of osteoclastogenesis by Salubrinal and Guanabenz

“Synthetic agents such as salubrinal and guanabenz, which attenuate stress to the endoplasmic reticulum, are reported to inhibit development of osteoclasts. However, the mechanism of their inhibitory action on osteoclasts is largely unknown. Using genome-wide expression profiles, we predicted key transcription factors that downregulated nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), a master transcription factor for osteoclastogenesis. Principal component analysis (PCA) predicted a list of transcription factors that were potentially responsible for reversing receptor activator of nuclear factor kappa-B ligand (RANKL)-driven stimulation of osteoclastogenesis. A partial silencing of NFATc1 allowed a selection of transcription factors that were likely to be located upstream of NFATc1. We validated the predicted transcription factors by focusing on two AP-1 transcription factors (c-Fos and JunB) using RAW264.7 pre-osteoclasts as well as primary bone marrow cells. As predicted, their mRNA and protein levels were elevated by RANKL, and the elevation was suppressed by salubrinal and guanabenz. A partial silencing of c-Fos or JunB by RNA interference decreased salubrinal- and guanabenz-driven reduction of NFATc1 as well as tartrate-resistant acid phosphatase (TRAP) mRNA. Collectively, a systems-biology approach allows the prediction of a RANKL-salubrinal/guanabenz-NFATc1 regulatory axis, and in vitro assays validate an involvement of AP-1 transcription factors in suppression of osteoclastogenesis.”

“Salubrinal and guanabenz are potent chemical agents for the inhibition of protein phosphatase 1 (PP1) that specifically de-phosphorylate eIF2α. Through upregulating the phosphorylated level of eIF2α and reducing translational efficiency of most proteins except for a limited set of proteins, such ATF4, these agents attenuate stress to the endoplasmic reticulum. Gene regulation by salubrinal and guanabenz, however, not only takes place at the level of translation but also at the level of transcription. In osteoclasts, it has been shown that administration of salubrinal and guanabenz suppresses receptor activator of nuclear factor kappa-B ligand (RANKL)-driven activation of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) ”

“A partial silencing of c-Fos and JunB decreased the mRNA and protein levels of NFATc1. Furthermore, there was a feedback loop in which a decrease in c-Fos by salubrinal reduced NFATc1 expression, and the reduction in NFATc1 further attenuated the level of c-Fos protein. AP-1 proteins are known to play a critical role in osteoclast differentiation. It is reported that mice lacking c-Fos are osteopetrotic due to abnormal development of osteoclasts”

Unfortunately, no real LSJL insights in this study.

Update #16 – More Resources – August 1st, 2014

Last month I could not write up a post for updates and the reason was because the website was down for more than a week. There was some technical and legal issues which needed to be resolved. Here are some things that has occurred in the last 2 months

• We’ve established that Dr. Ballock at the Cleveland Clinic has been working on a project to study on how to grow a fully functional growth plates with a colleague, a Dr. Eben Alsberg, who I have previously spoke very highly of. The project is supposed to be finished in a couple of months so there is going to be some big news that will be coming out soon.
• I’ve established that in Google Patents there are multiple types of patents which show that researchers have been working on the science of remodeling bones, using different chemicals.
• Regenexx and Mesoblasts have both been trying to win over in the spinal disc regeneration niche of biotechnology but their amazing lab animal study results might not be able to be translated to humans. This is the first real example that what we find in the lab may not work in real life.
• I have reported on the news of Alexander Teyplashin’s team and shown that they are dead serious on putting neoepiphyseal cartilage into human bones to let them grow again. This was the most shocking news to come out and it did lead to some talk in the small community.
• Teplayshin’s team filed multiple patents and papers showing how to start from human adipose tissue, get stem cells from them, turn them into chondrocytes, and deposit into scaffolds. They figured out the vascularization problem.
• It seems that many of the people I have mentioned have been working in collaboration for quite a while and I wasn’t aware of it until now. For example, Dr. Jean Welter of the Case Western Reserve University, which is based in Cleveland has collaborated/worked with Dr. Ballock, as well as Dr. Eben Alsberg. It shows that like almost any other micro-niche industry and field, all of the people doing their thing seems to know everyone else that is working on similar projects. I will be talking much more about Dr. Welter’s research soon though.
• I have purchased multiple books in the last two months looking deeper into the anatomy and physiology on how joints function. They include the following “Joint Structure & Function, A Comprehensive Analysis – 4th Edition” by Pamela K. Levangie and Cynthia C. Norkin, “Treatment by Manipulation” by AG Timbrell Fisher, and “The Complete Illustrated Book of Yoga” by Swami Vishnudevananda. Some purchases have been to help me and others around me to develop a much more active and healthier lifestyle.

As for myself, I have been going to an acupuncturist (who was trained in Russia with a Ph.D and M.D) so have gotten myself much more interested in some of the more quasi-scientific/alternative approaches to medicine and healing. In the last  month or so I have looked into some very strange forms of bone and joint treatment.

As for the backend of the website, I changed the ownership of multiple part. I stopped making any type of adsense earnings on the website – I changed all the adsense ID #s to be under Tyler’s name. There is going to be some big things that I planning on releasing in the coming half of this year.

Updates For Regenerative Tissue Engineering Using Stem Cell For New Growth Plates – How Much Longer?

So the most recent relevant post I had written had said that we would need to wait 30 or even 50 years before the limb lengthening surgery alternative would be finally be available. That made many people very upset that this idea which I had claimed to be “almost there” would still take that long. Let me try to give you guys a better estimate on what is reasonable to expect to happen within the 21st century in terms of biotech.

The short answer is this. If we were to be extremely optimistic, It would take 10 years for the stem cell technique to be viable for for humans to use, but only for real medical issues. Within 5 years, the physicians performing the technique would realize that there is a large financial incentive in using the stem cell approach on people for cosmetic reasons. So being very optimistic, it would take 15 years before we can finally have the chance to use the new growth plate method. On the opposite end, it would take maybe 30 years for the stem cells to have all the technical details worked out for them to be for use for humans, and still another 10 years before doctors are willing to put the technique for use to lengthen bones. So the worst we can do is wait 40 years. Yes, it sucks that it could take almost half a century, and maybe some of us might not even live that long, but biological science is not like computer science, where you have dedicated people who can pump out a new product in a dorm room with a computer working 24 hours a day.

Here is something else which we need to consider. Exponential Growth. When we remember Ray Kurzweil’s Law of Exponential Growth/Returns which is supposed to signal the point known as the Technological Singularity, it suggest that maybe, just maybe the technology may come even faster, if the exponential growth of technological advancement is true. That might mean that at the earliest, the idea of implanting new lab grown growth plates into bones to lengthen the bone might be available for at least young kids suffering real orthopedic limb angular deformities within as close as 5 years, and have it available for adults for cosmetic application within 10 years.

Let’s give you guys some perspective. If you guys typed into Google right now the term “new growth plate”, you would find as the first link a article/reference to Dr. Robert Ballock’s research at the Cleveland Medical Clinic. That shows that Ballock is very serious on getting the technical details on lab grown growth plates to work out. His desire is to use his knowledge from Harvard to help young children to correct bone grown irregularities. The article you find #1 in Google Search is called “Regrowing Growth Plates: A Fix for Kids’ Injuries? Promising research to mend growth-stifling plate damage” was written back in Feb of this year, 2014. The website by Cleveland Clinic showed that he is focused on the research, and his grant (which we linked to on the previous post) revealed that the project would end around August-Sept this year.

Quote… “In test tube studies, Dr. Ballock’s research team developed a 3-D cell culture model that replicates the key features of what appears to cause growth plate cartilage to develop in the column-like shape that leads to lengthening of the long bones in children.”

At the end, Ballock stated in typical research fashion that his research would be years away from being used for real application. This is disheartening to many teenagers and young people who want to be taller NOW and want something as soon as possible, but it takes time. Even when you want to use the stem cell approach for cosmetic use, other entities can resist you and your research.

The recent reveal that Alexander Teplyashin’s team in Mocow, Russia in getting bones to lengthen by implanting culture grown “growth plates” and trying them out on ram/sheep shows that in theory and concept, the stuff would work. However, Teplyashin is still a researcher (actually has a Ph. D in plastic surgery) and he must follow the guidelines and protocols that the academic and corporate rules set for him.

Here are some problems that have caused the whole stem cell research to be resisted. When George Bush was president (the 2nd one) he imposed laws which made it not possible for USA based researchers to use embryo derived stem cells. Because of Bush, the research into stem cells was halted for his entire 2 terms, which was basically a full decade. While the USA was held back, other countries in the world like Japan, South Korea, Germany, and Russia rushed forward and was willing to take the chance the Americans were not. It wasn’t until 2009, when Obama came to office did the law against stem cell research (at least the embryo derived ones) get lifted. (source: Obama Lifts Bush’s Strict Limits on Stem Cell Research).

The 2nd problem is that Teplyashin’s research has also been halted by the government branched in Russia, one source we found stated. His team is written up grants, petitions, and proposals to move forward in their research, but the government seems to be blocking his team from further progress.

All of this may sound like really bad news, but there is a LOT of good news which I have found recently which shows that the endeavor is definitely being pushed forward.

The Good News

Good News #1 – On the problem on cartilage material strength and whether it can handle the upper weight loading – It seems that this is not a problem. I do read a lot of reddit threads and on one reddit thread, where people were talking about the possibility of getting lab grown cartilage transplanted into their knees for pain management and osteoarthritis treatment ( http://www.reddit.com/r/science/comments/24gghv/for_the_very_first_time_engineers_have_grown/) it was revealed that cartilage that is created in the lab synthetically can withstand the weight load, with the high enough equilibrium compressive strength/modulus. Refer to the study Comparison of the equilibrium response of articular cartilage in unconfined compression, confined compression and indentation for more information.

Good News #2: The problem that the people who want to fix their knee articular cartilage degeneration have cleared up is that cartilage implants like autologous chondrocyte implantation has not worked too well to the defects on the surface of articular cartilage. The problem is that the neo-cartilage tissue doesn’t seem to be able to fused properly with the older, natural cartilage. The cartilage then turns into the fibrocartilage tissue type. Of course, these guys on the Reddit Science Thread wants to add cartilage to an area of the body where there is already cartilage. As we know, cartilage is an anareobic environment aka avascular, where the cells inside the ECM of the cartilage can’t get nutrients through blood vessels. Of course then it would make sense that plopping new cartilage on top would not work out too well, since traditionally you need vascularization to get the transplants to fuse with the rest of the body. Our desire is instead to add cartilage to a place where there is no more cartilage (fully fused) and only bone. Bones are very nicely vascularized. In fact, we reported news recently that the vascularization issue was figured out by Teplyashin’s team in a paper that they published (How Svetlana Zagirovna Sharifullina Developed A Vascularized Fully Functioning Growth Plate)

(Side Note: Here are some places which I regularly read up on and make sure I keep up with the most up to date information on reconstructive tissue engineering, stem cells, and such. http://www.reddit.com/r/science, http://www.reddit.com/r/stemcells, http://www.reddit.com/r/tissueengineering, http://www.reddit.com/r/Futurology/, http://www.reddit.com/r/orthopaedics, http://www.reddit.com/r/cancer . In addition, I also read up on the works for the dozens of researchers cited http://www.beckersspine.com/spine/item/14329-10-spine-surgeons-researching-disc-regeneration)

Conclusion:

So when it comes to the technical problems of getting the synthetic lab grown growth plates to meld with the surrounding bones, which is usually just a vascularization problem as Hakkers stated in our interview years ago, that seems to have been resolved.

And when you look at how strong the lab grown cartilage really are, looking at the values of the equilibirum compressive modulus, you find that the major issues that I have come up with seems to be almost all gone.

The few problem that I can think of is that the scaffold derived epiphyseal hyaline cartilage is not producing enough ECM organic compounds, most specifically Collagen Type II, the GAGs, and proteoglycan. However, I would suspect that the technical issue would be a rather easy one to figure out. The other thing is how would the surgeon make the right type of cut into the epiphysis-metaphyseal connected region to implant the cartilage. If you look at how real cartilage is layered in the human body (I saw off the end of a dead young pig’s leg), they are not flat, but curved and bumpy, which is probably due to the different rates of chondrocyte columnar activity.

Then when you look at how easily it is for people with a 3-D printer to print out layers of hydrogel and scaffolds these days, with maybe dozens of people doing similar research, things seem to be looking very good.

I don’t want to make it sound too positive, since there will always be huge issues. The article I found written by someone at Regenexx over the fact that Mesoblast spent hundreds of millions but still couldn’t get their human subjects to have similar results as their lab rats show that sometimes the best results on lab animals can not be translated to humans. The article back in May of 2013 MAJOR STUDY UPDATE: STEM CELLS EASE BACK PAIN revealed that while the subjects did indeed decrease back pain, the disc height did not increase like what happened in the lab with the rats.

Harald Oberlander who I have been in contact with referenced Mesoblast’s research to the people on the Make Me Taller forums but I don’t think even he is aware of the most up to date releases that is coming out of the company. Progress is not easy.

A Suggested Idea On How That We Can Theoretically “Reopen” The Closed Growth Plates

I have said multiple times that it would be next to impossible within this century to create any type of technology to ‘reopen’ fully closed growth plates, at least completely non-invasively. That has been my opinion for a long time since I really started to understand how human bones really work and are organized. Unless there is some type of radical biomedical technology I am not aware of that is coming down the pipeline in the next few decades, I would not be looking for a completely non-invasive way to get the growth plates back.

However, I did recently read an article which seems to suggest at a very theoretical level that it might be possible to kind of “reopen” the closed growth plates, but it would still not be completely non-invasive. Refer to the paper “Roles of neutrophil-mediated inflammatory response in the bony repair of injured growth plate cartilage in young rats“. The PDF for the article is free for download.

I was made aware of the paper when I was trying to find relevant research that Cory Xian had done, since Tyler said that Xian was one of those people we should be following. This paper I did take the time to really look over extensively, and it just slightly hints at the idea that we might be able to change the progenitor cell lineage to something which it is not naturally supposed to go into.

Here is what you need to understand about the microbiological processes that seems to occur after the growth plates in a rat model were intentionally fractured and then analyzed.

Process #1: Inflammatory – This would be the initial injury induced inflammatory response, which is the automatic process that is activated when almost every part of the body is injured. What happens is that inflammatory cells, which comes in the types known as neutrophils, monocytes, lymphocytes, starts being rushed to the local area of injury. In terms of time, at least on the rat models, the first process lasts around 3 days, the maximum effect being around 1 day.

• Process #2: Fibrogenic –
• Process #3: Osteogenic –
• Process #4: Bone Bridge Maturation –

More Details On the Initial Inflammatory Response – The researchers have identified at least 5 different types of chemicals which will be stimulated to go towards the injury area. They include a chemokine, pro-inflammatory cytokines, and fibrogenic growth factors.

• 1 type of neutrophil Chemotactic Chemokine – CINC-1
• 2 types of Pro-inflammatory Cytokine – TNF-Alpha and IL-1Beta
• 2 types of Fibrogenic Growth Factors – TGF-Beta1 and PDGF-B

What the researchers did was add a type of chemical which will decrease the number of neutrophil cell count. Quote: “Using an antirat neutrophil antiserum, a significant 60% reduction in neutro- phil count was achieved at the growth plate injury site”

(Side Note: The most interesting part of the neutrophil to pro-inflammatory cytokine stimulation is that while the antis-serum reduced the neutrophil count, it did not decrease the expression and numbers of the TNF-Alpha or the IL-1Beta (or the Fibrogenic Growth Factors like TGF-Beta1 and PDGF-B) . The reasoning is because besides the neutrophil expression increasing in the bone fracture site, there is two other types of cells which are rushed there, the monocytes and the lymphocytes, and the monocytes, which are also known as macrophage, are the other type of cells which really increase the expression of the Interleukin (IL-1Beta) and the tumor necrosis factor (TNF-Alpha). The results is that the number of MSCS which rushed to the local fracture sight was not not decreased. That suggested that the number of neutrophils in the growth plate injury sight had no real link to the amount of MSCs which will go to that growth plate injury area.)

The result of adding the chemical was that in the injury area, there was more bone tissue than cartilage tissue by ratio that had developed. This suggests that neutrophil cell types has the effect of at least shifting the osteogenic/chondrogenic ratio the other way. The researchers tested the osteogenic and chondrogenic gene expressors the SOX-9 and the COl-Type IIa and saw that the gene expressions were reduced. In reverse, the expression of bone cell differentiation transcription factor cbf-alpha1 and bone matrix protein osteocalcin, which create osteogenic tissue was increased.

If there is an injury aka fracture in the bones, the neutrophils would be able to help the fibrogenic growth factors to develop more cartilage progenitor type tissue and cells.

I quote the last sentence that is in the paper…