I went to this conference for two main reasons. The first was to talk with a company that was selling a 3D Bioprinter. The 2nd was to listen to the lectures and presentations by the leading edge researchers in this field to see if anything that they are working on can be applied to what we are doing here.

Here is what I can say.

Every single industry, field, or niche has a few people who become very important in their chosen area of expertise. From this one conference, I did find out who are the main people we should be focusing on and following.

• Dr. Lawrence (Larry) Bonassar
• Dr. Anthony Atala
• Dr. Warren Grayson

It turns out that a Dr. Jason Spector who was a speaker at the conference giving his talk  “Tissue Engineering, Bioprinting, and the “Reconstructive Ladder”” alluded to the fact that he was working with Dr. Bonassar on research with MSCs (Mesenchymal Stem Cells) to develop cartilage, whether it be fibrocartilage or hyaline cartilage. When I was listening to him, the name sounded very familiar until I realized that I saw the name before and actually wrote a big post on the work being done by Dr. Bonassar’s team in Cornell where they grew an implantable spinal disc. Refer to the post “This Researcher Succeeded In 3D-Printing Spinal Discs Allowing Adults With Closed Growth Plates To Grow Taller If They Desired – Big Breakthrough“. Dr Bonassar’s work and his Lab should be one of the primary focuses for us.

The other speaker that spoke about relevant information was a Dr. Paul Gatenholm (talk was “3D Bioprinting of Human Cartilage and Skin with Novel Bioink”, who apparently was a protege/student of Dr. Atala. He has a company CellLink where his son works at which has produced a type of bioink that is used in 3D Bioprinters. This bioink acts as the medium that stem cells would go into which is something similar to Hydrogel/Alginate/Extracellular Matrix/Scaffold. It is made from a derivative of cellulose.

Dr. Atala is sort of a super-star in the field of tissue engineering, regenerative medicine, and cartilage generation. He has been alluded to multiple times in the conference and I had referenced his research before as well on this website. Refer to the post ” Increase Height And Grow Taller Through Bioprinting And Electrospinning“. If you read this previous post, you would see that there was a Youtube video of Gabor Forgacs, who explained the revolution in regenerative medicine. He was the speaker at this recent conference which started everything off.

There was a third speaker Rahul Tare (talk was  “Application of Custom-Built Acousto-fluidic Perfusion Bioreactor for Cartilage Tissue Engineering” who revealed that he was also working with MSCs to engineer cartilage tissue.

It turns out that this conference I went to has people associated with the big players in this field. Most people there already knew each other.

In terms of the technical, I realized that bioprinting cartilage tissue that can be implanted back into the body is an endeavor which is probably impossible. In a discussion with Dr. Michael Gelinsky, he did not think that bioprinting a hyaline cartilage was viable at least for a long time. However, the idea of slowly developing chondrogenic tissue from implanted autologous chondrocytes and/or MSCs into a scaffold is probable, if not very doable. If Dr. Teplyashin’s group’s results are any indication, this step has already been done.

The problem which the researchers were trying to solve was over vascularization. Vascularization was the main problem that people who want to bioprint organs need to figure out. It was the bottleneck.

The other problem is over regulations. It turns out that many of the most scientifically advanced countries who are working in tissue engineering and regenerative medicine will not allow organs be implanted into the patients body.

Here are the list of 3D Bioprinters that I found out about.

1. BioBots (3d Bioprinter)
2. RegenHu 3d Bioprinter
3. EnvisionTec Bioprinter
4. Organovo’s 3D Printer
5. Regenovo (China based)
6. Qingdao Unique Products 3D Bioprinter
7. Izumi International Inc (Deposition Machine turned into 3D Printer)

Some of the models were cheap but some were extraordinarily expensive.

Some others things I realized that I made a mistake on was to forget about the step of bioreactor. You need a bioreactor for the implanted cells into a culture or scaffold, with the growth factors, to proliferate in numbers. Dr. Spector revealed that to get the MSCs to differentiated into a tissue that is actually worth something, you need at least 200 million cells to work with. Anything less and the tissue that has been formed is not really enough.

So to make a correction on the steps, it would be

• 1. Biopsy of bone marrow to get MSCs/ Extract part of the iliac crest for chondrocytes
• 2. Use collagenase to dissolve the ECM around your desired cells
• 3. Spin the solution to separate the cells from the other compounds
• 4. Get the right type of cell medium (alginate, hydrogel, etc.) to put the cells in.
• 5. Instead of medium, it could be a scaffold shaped in whatever way you want.
• 6. Add some type of growth factor into the scaffold to help the cells differentiate or proliferate.
• 7. Put it into a bioreactor to make the cells proliferate.
• 8. Induce vascularization in the tissue (this is the tough part)
• 9. Take the scaffold/medium out and implant the grown  tissue back into the person’s body. Wait until vascularization occurs to connect the new implant with the rest of the patient’s body.

Notice how I did not say bioprinting anywhere. Remember that there is more than 1 way to get the stem cells/chondrocyte implantation to work.

There is actually 2 ways to do this.

1. You can use a 3D Bioprinter to bioprint a fully functional growth plate, with the chondrocytes inside in columnar structure form
2. You can put extracted MSCs or chondrocytes into a scaffold (hard structure) and grow it into a bioreactor. You eventually implant the scaffold between the bones and wait for the scaffold to slowly turn into a hyaline cartilage layer of tissue ala pseudo-epiphyseal plate cartilage.

The 1st way is going to be very difficult, but not impossible. The 2nd way is not as “clean” but it should work.

There will be more conferences in the coming months worth looking into. They are….

• 2015 4th Termis Tissue Engineering World Conference – Dr. Atala will be a keynote speaker there.
• Biomaterials & Tissue Engineering Gordon Research Conference – Dr. Warren Grayson will be a speaker there.
• Tissue Engineering, Synthetic Biology & Bioprinting (2016) – by SelectBio
• Innovations in Cell-Based Regenerative Therapies Conference (MSC 2015) –

What you sort of realize as you go down the list of speakers and attendees is that the same people was at this conference too. Eventually after so many conferences you meet the same 300 people over and over again.

 

 

Prx1 seems to be a very promising target for height increase as it seems to have targets early in development.  Unfortunately, I couldn’t find any Prx1 stimulating substances but hopefully you can?  Srx1 is involved in the repair of Prx1 so that could be another target as well.

Regulatory divergence modifies limb length between mammals

“Natural selection acts on variation within populations, resulting in modified organ morphology, physiology, and ultimately the formation of new species. Although variation in orthologous proteins can contribute to these modifications, differences in DNA sequences regulating gene expression may be a primary source of variation. We replaced a limb-specific transcriptional enhancer of the mouse Prx1 locus with the orthologous sequence from a bat. Prx1 expression directed by the bat enhancer results in elevated transcript levels in developing forelimb bones and forelimbs that are significantly longer than controls because of endochondral bone formation alterations. Surprisingly, deletion of the mouse Prx1 limb enhancer results in normal forelimb length and Prx1 expression, revealing regulatory redundancy. These findings suggest that mutations accumulating in pre-existing noncoding regulatory sequences within a population are a source of variation for the evolution of morphological differences between species and that cis-regulatory redundancy may facilitate accumulation of such mutations.”

“One developmental control gene known to promote limb skeletal elongation is Prx1, also called MHox or Prrx1. Prx1 is a paired-related homeobox gene expressed in somites, craniofacial mesenchyme, and limb mesoderm during mouse development”

“the forelimbs of Prx1BatE/BatE mutants are on average ∼6% longer than their wild-type littermates at E18.5 ”

“The forelimbs of Prx1-null homozygotes are ∼12.5% shorter than controls at E18.5”

“average mutant long bones express ∼70% more Prx1 than wild-type siblings”

“Long bone chondrocyte proliferation is elevated by ∼6% at E15.5 in Prx1BatE/BatE mutants.”

“[The Prx1 induced] limb elongation arises at stages of mouse gestation when Prx1 expression is limited to the perichondrium of the developing limb skeletal elements. It is known that the perichondrium is an important regulator of endochondrial bone growth”<-Could this be related to the zone of Ranvier which should be connected to the perichondrium?  Since the zone of Ranvier is linked to an earlier developmental state of the growth plate it could mean that Prx1 could be helpful in inducing neo growth plates.

According to this grant, Prx1 inhibits bone formation so it favors an earlier developmental state overall.

Prx1-Expressing Progenitor Primary Cilia Mediate Bone Formation in response to Mechanical Loading in Mice

“Increases in mechanical loading can enhance the addition of new bone, altering geometry and density such that bones better withstand higher forces. Bone-forming osteoblasts have long been thought to originate from progenitors, but the exact source is yet to be identified. Previous studies indicate osteogenic precursors arise from Prx1-expressing progenitors during embryonic development and adult fracture repair. However, it is unknown whether this cell population is also a source for mechanically induced active osteoblasts. We first identified that Prx1 is expressed in skeletally mature mouse periosteum, a thin tissue covering the surface of the bone that is rich in osteoprogenitors. We then traced Prx1 progenitor lineage using a transgenic mouse model carrying both a Prx1-driven tamoxifen-inducible Cre and a ROSA-driven lacZ reporter gene. Cells that expressed Prx1 when compressive axial loading was applied were detected within the cortical bone days after stimulation, indicating osteocytes are of Prx1-expressing cell origin. In addition, we evaluated how these cells sense and respond to physical stimulation in vivo by disrupting their primary cilia, which are antenna-like sensory organelles known to enhance mechanical and chemical signaling kinetics. Although Prx1-driven primary cilium disruption did not affect osteoblast recruitment to the bone surface, the relative mineral apposition and bone formation rates were decreased by 53% and 34%, respectively. Thus, this cell population contributes to load-induced bone formation, and primary cilia are needed for a complete response. Interestingly, Prx1-expressing progenitors are easily extracted from periosteum and are perhaps an attractive alternative to marrow stem cells for bone tissue regeneration strategies.”

“periosteum, which surrounds bones and is rich in progenitor cells known to preferentially differentiate towards the osteogenic lineage”

“physical stimulation activates and encourages osteogenic differentiation of progenitors within the periosteum.”

“One potential mechanism by which progenitor cells may become mechanically activated is through the primary cilium. Primary cilia are antenna-like organelles that extend from the cell surface and serve as signaling microdomains.”

“osteogenic response to fluid shear is lost when periosteal progenitor primary cilia are disrupted in vitro”

“Prx1-expressing cells become embedded osteocytes in response to physical loading and this mechanism requires the primary cilium.”<-if Prx1 enhances longitudinal bone growth and Prx1 mainly effects on chondrogenic cells maybe there are non chondrogenic ways to enhance longitudinal bone growth.

It seems that there was another legitimate program that was developed back in the early 90s by a Russian doctor who claimed that he could help people stretch their back and body to make them taller. It is very similar to the A-Grow-Bics program that came out by the French Trainer a few years back. There was even a few interviews and media coverage that came out with his claims. Based on what Michael Goldreyer (Goldreer) said, the clinic seemed to have gone bankrupt very quickly after it opened up back in the early 90s. However it is still interesting to see that the rumors that was coming out of russia of a doctor who developed a program which the book “School of Height” (PDF available for free download in the free section) was based on real events.

This will be the first of maybe a dozen videos that will be uploaded, almost all in Russian (a few will be in armenian and arabic). I plan to release the rest over time in the coming months.

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One of the readers of this website informed me about the progress of another company called EpiBone which has been involved in growing bones that can be implanted back into the body with a high chance of complete functionality.

At first when I looked over the website and what the company was claiming, it looked almost exactly like the Israeli Biomedical Company Bonus Biogroup which I had written a post about before (Here).

Again, the problem is not that bone grafts can’t be built from scratch. They can, and it is very easy. The problem is when you try to build a bone-cartilage structure which functions. Building a functional cartilage structure has always been the difficult part. Even harder is to grow or build a bone-to-cartilage structure which operate together as one.

I was just about to write off this company as another company which will not push further until I looked at the list of authors who wrote the original scientific paper “Engineering custom-designed osteochondral tissue grafts”

The first name that is associated with the paper is what got me excited. It is Warren Grayson!!

I had said before from many posts written back in 2013 and 2014 that Dr. Grayson’s research is one of the few that is really going to get us to a Limb Lengthening Alternative. If Dr. Grayson is associated with EpiBone, as say the lead Research Scientist, then I would suspect that we maybe 5 years ahead of schedule in getting the Tissue Engineering approach to be successful. Where as I had predicted as long as 20-30 years before, I have to reassess that time line down to even 15 years.

However, when you look at the team that is actually listed on the website only one name is also found associated with the paper, a Gordana Vunjak-Novakovic. A Sarindr Bhumiratana is the other name that is worth looking at since he seems to be an expert on tissue engineering. There was a similar paper written to a 2nd source “Engineering anatomically shaped human bone grafts”

This looks like the real deal guys. They know that growing bones to be implanted back into the body is easy, and already done by other research teams and companies around the world.

It comes down to whether they can put enough research, energy, and funding to get the Bone-Cartilage type of tissue graft to work out which is what is going to determine whether we will see something that is available for the regular patient/customer within the next 10-15 years.

Apparently there has been a lot of media and press surrounding this company with some big name newspapers and online content aggregators talking about this company up. Bloomberg, CNN, Forbes, The Guardian, etc. are all getting some information on this company.

Cross your fingers guys. I will be going to the Organ-On-A-Chip BioMedical Conference/ 3D Bioprinting Conference in Boston (on Beacon Hill) from July 7-9, and then write a long post to report back to you guys telling everything that I have gathered about the most cutting edge research that is coming out today from the labs of Universities. I will mention this company EpiBone to the professors there and see if they are also working on the same thing or have maybe even surpassed what we are seeing with this company. I need to remind the readers that I paid $1,700 of my own money to get into this conference, since I am this dedicated to the cause. When I say I am going to put my money in on this endeavor, I have.

In addition, I recently also purchased the book “Mesenchymal Stem Cells and Skeletal Regeneration” off of Amazon and plan to read up on it on the plane to make sure that my level of understanding of the tissue engineering techniques are at the level of the presenters and exhibitors at the Conference. I might even record the conversations I have to upload it to the website for all of you to hear.

PS: I will be posting up quite a few videos to the Natural Height Growth Youtube Channel over the next few months of videos mostly in Russian of height increasing programs. Some of them show Alexander Teyplyashin’s research, and even AS Palko’s School of Height program in action, as well as old interviews done in russian of people who went through with LLS.

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