Tuesday, May 31, 2011

Grow Taller with Aqua Pressure?

Previously, I dismissed Acupuncture(and Aqua Pressure) to increase height.  The videos by Dr. Prakash Verekar did not fully prove height growth and that the height growth may be temporary height due to muscle relaxation or laying down allowing for replenishment of the proteoglycans in the spine.  However, we have learned of the important of water in inducing differentiation of stem cells.  LSJL is based on inducing hydrostatic pressure in the epiphysis.

If water is so important to our theories then couldn't injecting water directly into the epiphysis and cartilage increase hydrostatic pressure?  Well water may diffuse directly immediately but it would still temporarily cause shear strain and hydrostatic pressure.  It would also have to be tested in animals for safety but it should be easier and more cost effective.  All you'd need is a syringe, water, and a Sprague-Dawley rat(plus feeding supplies).  Test out various injection possibilities.  If you don't want to sacrifice the rat you can always just measure it alive.  Just drinking more water wouldn't work because you would be reliant on where your digestive system sends water to.

  In trying to ask scientists about exactly how endochondral ossification increases height, I received an interesting comment from Farshid Guilak.  If endochondral ossification was a merely transformative process from cartilage to bone then limbs would stay the same size!  There must be some part of the process that causes an increase in bone size.  There are two possibilities.  One could be due to the negative charge of the cartilagenous matrix which when turns to a no charge generates force. Sort of like the electron transport chain or the kreb's cycle.  Water is still likely key in this process as the combination of 2 Hydrogens and an Oxygen generates potential energy that can then induce bone formation.

Farshid Guilak presented this possibility: "since cartilage has a lot of water and bone doesn't, so there is the possibility that it could expand durring ossification, or simply replace water and other components with mineral."

With aqua pressure you are changing the typical balance of water in bone which could lead to an adaptation that increases height.

Here's a paper that shows one instance of hydrolysis during endochondral ossification:

Characterization of the phosphatidylinositol-specific phospholipase C-released form of rat osseous plate alkaline phosphatase and its possible significance on endochondral ossification.

"Alkaline phosphatase activity was released up to 100% from the membrane by incubating the rat osseous plate membrane-bound enzyme with phosphatidylinositol-specific phospholipase C[we'll call this PSPC]. The molecular weight of the released enzyme was 145,000 on Sephacryl S-300 gel filtration and 66,000 on PAGE-SDS, suggesting a dimeric structure. Solubilization of the membrane-bound enzyme with phospholipase C did not destroy its ability to hydrolyse PNPP, ATP and pyrophosphate. The hydrolysis of ATP and PNPP by phosphatidylinositol-specific phospholipase C-released enzyme exhibited 'Michaelian' kinetics with K0.5 = 70 and 979 microM, respectively. For pyrophosphate, K0.5 was 128 microM and site-site interactions were observed (n = 1.4). Magnesium ions were stimulatory (K0.5 = 1.5 mM) and zinc ions were a powerful noncompetitive inhibitor (Ki = 6.2 microM) of phosphatidylinositol-specific phospholipase C-released enzyme. Phosphatidylinositol-specific phospholipase C-released alkaline phosphatase was relatively stable at 40 degrees C. However, with increasing temperature from 40-60 degrees C, the enzyme was inactivated rapidly following first order kinetics and thermal inactivation constants varied from 5.08 x 10(-4) min-1 to 0.684 min-1. Treatment of phosphatydilinositol-specific phospholipase C-released alkaline phosphatase with Chellex 100 depleted to 5% its original PNPPase activity. Magnesium (K0.5 = 29.5 microM), manganese (K0.5 = 5 microM) and cobalt ions (K0.5 = 10.1 microM) restored the activity of Chelex-treated enzyme, demonstrating its metalloenzyme nature. The stimulation of Chelex-treated enzyme by calcium ions (K0.5 = 653 microM) was less effective (only 26%) and occurred with site-site interactions (n = 0.7). Zinc ions had no stimulatory effects. The possibility that the soluble form of the enzyme, detected during endochondral ossification, would arise by the hydrolysis of the Pl-anchored form of osseous plate alkaline phosphatase is discussed."

"a membrane-bound and a soluble form of rat osseous plate alkaline phosphatase appear during the ectopic mineralization process"

"soluble enzyme present during endochondral ossification could arise through the hydrolysis of Pl-anchored form of the enzyme by specific anchor-cleaving activities as was reported for liver alkaline phosphatase"

So the osseus plate membrane bound enzyme may become soluble by hydrolysis related properties during endochondral ossification.  So this enzyme may be key to how the changing of bone to cartilage increases height growth.  Hydrolysis(water reactions) are key to making this enzyme soluble and active.

Here's a study that shows the ability of water to induce endochondral ossification:

The in vitro chondrogenic response of limb-bud mesenchyme to a water-soluble fraction prepared from demineralized bone matrix.

"Demineralized adult bone matrix initiates de novo ectopic endochondral ossification 2-3 weeks following its intramuscular implantation into adult animals{this is injection directly into the muscle}. This phenomenon appears to be similar, in some ways, to inductive cell-matrix interactions which regulate cartilage and bone formation during development. In the present study, we used embryonic chick limb-bud mesenchymal-cell cultures to bioassay extracts of demineralized bone matrix for chondrogenic activity. Guanidinium-chloride (4 M) extracts of demineralized bovine bone were dialyzed against buffers of decreasing ionic strength and then cold water. The cold-water-soluble fraction was found to stimulate chondrogenesis in intermediate-density limb-bud cell cultures (2.2 X 10(6) cells per 35-mm dish), as revealed by visual inspection. Further fractionation of this material by anion-exchange, carbohydrate-affinity, and molecular-sieve chromotography produced a semipurified preparation possessing chondrogenic-stimulating activity at doses ranging from 3 to 10 micrograms/ml. The in vitro chondrogenic response of limb-bud mesenchymal cells was dose-dependent, required a minimal initial plating density of 2.08 X 10(5) cells/mm2 of culture dish, and developed gradually over 8-10 days. At an optimal dose of extract, a continuous exposure period of at least 2-3 days was necessary to produce detectable chondrogenic stimulation. In addition, the amount of cartilage formed following an 8-day exposure was markedly influenced by the culture 'age' of the mesenchymal cells (i.e., the time between plating and the start of treatment)."

Cartilage has water and bone doesn't.  The substance that was water soluble was what was effective in inducing chondrogenesis.  Water and hydrolysis are key distinctions between cartilage and bone.  The water levels might influence the microenvironment to induce chondrogenesis.  This is what we are trying to alter with LSJL the water microenvironment.

"adult bone contains substances that stimulate or control the process of  bone repair."

Here's something that explains how water is involved in the change of cartilage to bone:

Mechanisms of cartilage precursor replacement by bone in the mammalian skeleton.

"The initiation of the endochondral process in ontogenesis is preceded by structural and metabolic preparation of the cells of cartilage anlages of the skeleton as well as the satellite cells of the vascular terminations[likely VEGF related] adjoining the cartilage. In the cartilage anlage, due to a progressive specific biosynthesis, the macromolecular chondroitin-protein complexes capable of binding a large amount of water, are accumulated{both chondroitin levels and water levels may affect how much is stored}. As a result the volume of a swelling chondrocyte increases twentyfold or more and a high hydrostatic tension is created within it{the chondrocyte streches due to the water within it}. At the same time the metabolic situation within the cartilage favours a local concentration, near the hypertrophic chondrocytes, of the perivascular cells possessing an increased resorptive function. The resorption of the cartilage partition walls by these cells leads to the opening of swollen chondrocytes[the opening of these chondrocytes could result in water release that push bone apart]. The blood cells and the cells of the blood capillaries themselves are engulfed through the resulting crater into the emptied lacuna. In the lacuna the conditions of stasis are created which promote local redistribution of the supplied cells and reutilization of the cartilage destruction products. The perivascular cells concentrated along the remnants of the cartilaginous matrix are gradually differentiated into preosteoblasts and osteoblasts. The endosteal bony laminae are built on the remnants of the cartilaginous matrix. The perivascular cells of the apical zone of a lacuna retain their clastic function and continue spreading of the endochondral process on the adjacent chondrocytes. In the course of the successive destruction of hypertrophic chondrocytes, the system of vessels is formed in the endochondral centre, according to the sinusoid pattern."

Couldn't get this full study.

So it's likely the release of water by swollen chondrocytes after the reabsorption of tissues near the cartilage that pushes the bones apart and results in a larger size.  You'd need at least a large enough growth plate to build the matrix so they wouldn't swell right away.  Then you'd need sufficient force generated by the release of water to deform(stretch) the bone.

Abundance of evidence points to a huge role in water in endochondral ossification and differentiation of stem cells into chondrocytes.  Therefore it is possible for Aqua Pressure to work.

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