Monday, February 6, 2012

Growing Taller with a Bioengineered Cartilage Pellet

Growth plate transplants have shown signs of being successful.  A growth plate transplant is really nothing more than a layer of chondrocytes within bone which is what we're trying to induce with hydrostatic pressure LSJL.  A pellet usually refers to something of a very small size so likely utilizing bioengineered cartilage pellets would be much easier than limb lengthening surgery.

Restoration of longitudinal growth by bioengineered cartilage pellet in physeal injury is not affected by low intensity pulsed ultrasound

"We investigated the application of a Bioengineered Cartilage Pellet (BCP) in repairing a rabbit physeal fracture model, and the possible effects of Low Intensity Pulsed Ultrasound (LIPUS) treatment. Rabbits with physeal fracture created were assigned to the NC group (no BCP, no LIPUS), GC group (BCP, no LIPUS), and GT group (BCP and LIPUS). Femoral lengths and cartilage area were assessed at 4, 8, and 16 weeks post-defect. After transplantation, the BCP showed continuous growth in the host and demonstrated resemblance to a natural growth plate. The GC group showed 34.1, 32.1, and 41.1% advantage in lengthening over the NC group and the GT group showed 51.1, 41.6, and 26.9% improved lengthening than the NC group, at 4 (p = 0.203), 8 (p = 0.543) and 16 weeks (p = 0.049), respectively. Cartilage area was shown to be significantly higher in GC and GT group compared to NC group (p < 0.05). No significant difference was found between GC and GT group. Femoral longitudinal growth was shown to be improved by the BCP, however no additional enhancement effect was shown to be provided by LIPUS[maybe they targeted LIPUS on the wrong cells or the LIPUS stimulation wasn't enough?]. "

"mesenchymal stem cells (MSC) [can be used] together with a composite scaffold to regenerate cartilage [and it's possible to] use the cultured autologous chondrocytes together with a demineralized bone matrix to prevent bone bridging and restoring growth in damaged growth plate in rabbit tibia. "<-We've seen the importance of a demineralized bone matrix before.  And it's possible that a demineralized bone matrix can increase height without the presence of external stem cells or chondrocytes.

"[LIPUS has been] shown to increase chondrocyte proliferation, proteoglycan synthesis, and aggrecan gene expression."<-Which is why LIPUS may help increase height.

"LIPUS treatment started 1 day after introduction of the physeal defect and each rabbit in the GT group received 20 min of treatment, 5 days per week until time-points were reached. Coupling gel was applied to the ultrasound transducer with air bubbles carefully removed as they would block transmission of the ultrasound waves. The transducer was then placed directly on the medial side of the defective knee. The LIPUS devices delivered 200-μs bursts of 1.5 MHz ultrasound signals repeated at 1.0 kHz with intensity of 30 mW m−2."<-So the LIPUS device was applied laterally to the knee which is how we would apply LIPUS to induce stem cell differentiation into chondrocytes.  Maybe the fracture inhibited LIPUS effectiveness as the fracture may reduce hydrostatic pressure.  A study involving drilling done by the LSJL scientists showed that holes can reduce hydrostatic pressure.

"Our findings suggest skeptics that LIPUS may accelerate endochondral ossification in physeal fractures, which will cause premature physeal plate closure that is not always desirable in physeal injuries."<-So LIPUS may be bad for height growth during development?

"The lack of efficacy of LIPUS in stimulating the BCP in its transplantation site could be due to variations in ultrasound transmission to intact and fractured bone via an entry point. The bony ends of the physeal defect might have deflected off some of the ultrasound waves. This may be related to others' observation in the enhancement of fracture healing by LIPUS, but no positive stimulatory effect on bone mineral density in intact bone."<-So it's possible that LIPUS might not go deep enough into the bone to increase hydrostatic pressure and induce chondrogenesis.

Here's the study that describes how pellet culture was performed unfortunately I don't have access to it:

Bioengineering and Characterization of Physeal Transplant with Physeal Reconstruction Potential

"In this study, we attempted to treat proximal tibial physeal damage in rabbits with transplanted bioengineered physeal tissue. Resting chondrocytes from the reserve zone of costal cartilages of 6-week-old rabbits were pellet cultured in centrifuge tubes. The pellets were characterized histologically and biochemically with reference to the normal physis. The bioengineered tissue was then transplanted into partially damaged proximal tibial physis. Histological changes and proteoglycan metabolism of the transplants were monitored until 7 weeks posttransplantation. Our results showed that chondrocytes cultured by three-dimensional pellet exhibited cell division and the derived cells arranged in short columns similar to normal physis. They synthesized and deposited cartilaginous matrix and differentiated into hypertrophic chondrocytes marked by increases in cell size and alkaline phosphatase activity. The transplant incorporated well in host tissue with no sign of rejection for up to 7 weeks posttransplantation. A further 3-fold increase in thickness of the transplant within the host was observed. Endochondral ossification was demonstrated at 7 weeks posttransplantation."

Here's another study however that explained the method to form chondrocyte pellets:

Hyaline cartilage engineered by chondrocytes in pellet culture: histological, immunohistochemical and ultrastructural analysis in comparison with cartilage explants

"Fertilized White Leghorn chicken eggs were incubated at 37 °C for 16 days. The distal part of the sternum was removed from chick embryos, predigested in 0.2% collagenase for 30 min, and further digested in fresh collagenase solution for 3 h[likely to get red of the extracellular matrix]. Chondrocytes were suspended in supplemented Dulbecco's modified Eagle medium  at a concentration of 107 mL−1. Four × 106 chondrocytes in 0.4 mL of cell suspension were transferred into each 0.75-mL tube . Chondrocyte pellets were formed by centrifugation at 500 g for 10 min[so they use centrifugal force to get all the chondrocytes at the end of the test tube]. The culture medium was DMEM plus 10% fetal bovine serum, 50 µg mL−1 ascorbate, 2 mm glutamine and 0.2% penicillin/streptomycin. The pellets were cultured at 37 °C under a gas mixture of 95% air/5% CO2. The pellets were transferred into Petri dishes at day 3 and continued to grow for 2 weeks."

In the bone marrow there are no chondrocytes and thus no cartilagenous matrix to digest.  The medium is mostly things that already exist in the bone marrow.  What we can do is use centrifugal force to get more mesenchymal stem cells in one location.  The development of cartilage has been shown to be initiated by a condensation of mesenchymal precursor cells.  Maybe Kojima's rotation device can help you grow taller...  Kojima's green light could demineralize a bone surface and then the rotation could get a bunch of stem cells onto that bone surface.

So maybe after LSJL you need to use a bicycle to get stem cells onto a bone surface which has been demineralized by hydrostatic pressure.

13 comments:

  1. oh i cant wait to grow already im getting so impateint !
    the work you are doing tyler is truly amazing, if we do actually get a method to grow!
    i know you are a very patient person since you have to examine multiple growth mechanism and studies (like a scientist( lol but I think its time we started moving from theory to action! this would be truly amazing if we do indeed get a alternative to reactivating or getting new active growth plates, if i were there i would squeeze you so hard with all my thanks!! and truly it is a gift, so lucky to be alive at this time right now. so thankfull to have this information, thanyou for all this blogging, and lets get doing!

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    1. I agree, if there has been enough new information released on LSJL, I think a new routine should be set in place for people to try. I think there are still people trying the old method and not getting much (if any) results.

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  2. ok anom 1 i get your point but stop ass lickin will ya

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  3. "I think there are still people trying the old method and not getting much (if any) results."

    Is there a "NEW" method?

    And does tyler make summaries of the knowledge of each journal article and update it into a "general LSJL knowledge"-file?

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  4. where can i get that green light?

    i think its about time we start moving away from theoretical and into practical applied and proven.

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  5. If you want establish a new method then please help record data. Do LSJL, record additional stimulus you performed, and record any bone phenotype changes not just height.

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    1. I agree Tyler, but what is the new method we should be trying? I am asking you, not because you owe anyone anything, but because I trust your knowledge and believe through your finding (which I read but barely grasp) it seems there is a lot of new information out there than what was had when the first 1 minute clamp routine was set in place.

      The internet is a really powerful tool where great minds can come together to solve a problem. This is the kind of information that should be out there being discussed in an open format, molded, debated, whatever it takes to get the job done.

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    2. I've been doing LSJL since August. I grew a half inch from the beginning of September to Mid-sept. Then I grew another half inch at the end of the month (5'11 morning height). I'm 20 years old. Since Oct. I have plateaued, but I still do LSJL 5 times a week before bed. I also take Hyulronic Acid, B-complex, Melatonin (before bed), Niacin (every 3rd day 300 mg), Huperzine A 200 mg, and Glucotamine & MSM with a Emergen-C pack. This is my routine for LSJL: 1 minute load on lower femur, 1 minute load on upper tibia, 1 minute load on ankles/ lower tibia. I increase the load every 2 sec. like you said with 1/6th of a turn on the C-clamp. I tend to feel hydrostatic pressure more/better on my right leg than left (almost feels like cold fluid is flowing through your bones). Then I'll tap 30 times with the C-clamp on both ankles, and on both sides on both knees. Then I'll tap 30 times on both hips. Then I'll tap both femurs back and forth , and then I'll do both tibias and feet up and down. Then I'll tap the wrist, elbow, and shoulder 30 times on both arms. I have noticed my knees have gotten a little wider esp. my left, but I Feel I have a slight challenge loading the epiphysis (lower femur, upper tibia) on my left leg sometimes. I'll send you pics

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    3. I thought I read a few posts ago that said 3 mins of clamping may be best?

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  6. So regarding the centripetal force required to push the stem cells...

    Would that explain why basketball supposedly helps make you taller? Or even jumping in general for that matter?

    When you jump and fall from the apex of your jump, wouldn't the acceleration downwards help force the synovial fluid to go where it would most benefit?

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    Hypothetically speaking, could we do LSJL to demineralize then proceed to jump/kick our legs to augment the effects?

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  7. Tyler , does lipus by itself have any hope of making people grow taller

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  8. May be useful exogen 4000+?

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  9. You've said before that Kojima uses a "green light" to demineralize bone. where does it mention on their website? Has anyone been there before and confirmed this?

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