Grow Taller by manipulating the OPG/RANKL gradient

In our study of the bone multicellular unit(the osteon), we learned that there are RANKL and OPG fields which affect where bone is absorbed and where bone is deposited.  If you could say alter the OPG fields so that they are on the top and bottom of the bone then you can grow taller that way.

There is also evidence that osteoblasts can exist outside of an oteon unit and these osteoblasts are known as surface osteoblasts.  We already know that osteoblasts can deposit new bone lengthwise as that is an important part of endochondral ossification.

We know that Calcitonin alters the RANKL/OPG gradient.  Calcitonin increases the OPG(bone forming) mRNA in osteoblasts while lowering the RANKL(bone absorbing) mRNA in osteoclasts.  FGF-8a also altered the OPG/RANKL gradient by inhibiting RANKL.  Zinc deficiency alters the OPG/RANKL gradient towards RANKL.  Icariin alters the OPG/RANKL gradient towards OPG.  Runx2 via the DNA Methylation status of RANKL affects when RANKL is expressed during endochondral ossification(at the end in the hypertrophic phase).  In other words, Runx2 helps form a new OPG/RANKL gradient in the growth plates.  Curculigo Orchioides increased serum OPG levels.

What else can we learn about the OPG/RANKL gradient and how can that knowledge help us grow taller?


Bone lesions in multiple myeloma--the OPG/RANK-ligand system.

"Multiple myeloma[myeloma involves the mutation of white blood cells which causes alterations of the bone marrow] has recently been found to induce considerable imbalance in the newly identified system of osteoprotegerin (OPG), receptor activator of nuclear factor KB ligand (RANKL) and RANK[White blood cells have the ability to alter the OPG/RANKL gradient]. The binding of RANKL to RANK on the surface of osteoclastic precursors in the presence of m-CSF activates the signalling pathways for differentiation and proliferation of an osteoclastic line. OPG is a decoy circulating receptor for RANKL which blocks its binding to RANK. There are at least three mechanisms by which myeloma cells affects the OPG/ RANKL/RANK system: 1: The adhesion between the myeloma / stromal cells and the osteoblastic precursors stimulates the system by increasing the production of RANKL[so blood cells have the ability to stimulate osteoblast precursors]. 2: Some myeloma lines produce independently membrane-bound or free RANKL. 3: The normal and mutated plasma cells bind, degrade and block the OPG production from the stromal cells. The OPG/RANKL/RANK system is the latest therapeutic target in the treatment of myeloma bone disease. The first results from the application of a synthetic analogue of OPG, as well as of RANKL antagonists or RANK inhibitors show decrease of the number of osteoclasts, osteolytic lesions and M-gradient."

So if you alter the blood cells, you have the ability to alter where bone is deposited an absorbed.  The RANKL/OPG gradient is essential to any height increase method as it controls exactly where bone is deposited and absorbed.

PTH(1-34)[Teriparatide]-induced changes in RANKL and OPG expression by human PDL cells modify osteoclast biology in a co-culture model with RAW 264.7 cells

"Parathyroid hormone (PTH) is widely accepted as an anabolic agent when administered intermittently. Here, we explored the influence of intermittent PTH(1-34) on the expression of local factors by human periodontal ligament (PDL) cells that modify osteoclast biology. In a co-culture model of mature PDL cells and RAW 264.7 cells, intermittent PTH(1-34) induced an increased gene expression for tartrate-resistant acid phosphatase[alsoknown as TRAP] (+84%), cathepsin K (+56%), and vitronectin-receptor (+56%); and an enhanced resorptive activity of differentiated osteoclasts (+154%). These findings were correlated with a reduction of the osteoprotegerin (OPG)/receptor activator of nuclear factor kappaB ligand (RANKL) ratio in the presence of PTH(1-34; -44%)[. Similar results were obtained when RAW cells were cultured with the conditioned medium of PTH(1-34)-stimulated PDL cells. In contrast, when less mature PDL cells were co-cultured with RAW cells, PTH(1-34) induced an inhibition of osteoclastic differentiation (TRAP, -35%; cathepsin K, -28%; vitronectin-receptor, -35%), a reduction of the resorbed substrate area (-77%) and an increase of the OPG/RANKL ratio (+11%). The conditioned medium of PTH(1-34)-pretreated less mature PDL cells led to a down-regulation of the number and activity of multinucleated cells. These data indicate that intermittent PTH(1-34) modifies the expression of membrane-bound and secreted factors by PDL cells which then in turn alter osteoclast biology."

The effect of Parathyroid Hormone on the OPG/RANKL gradient is based on the cellular maturity.  So factors that effect this cell maturity may be more important than PTH itself.

Subchondral bone microstructural damage by increased remodelling aggravates experimental osteoarthritis preceded by osteoporosis.

"Accordingly, we assessed whether microstructure impairment at subchondral bone aggravates cartilage damage in [an] experimental model. OP[Osteoperosis] was induced in 20 female rabbits, by ovariectomy and intramuscular injections of methylprednisolone hemisuccinate for four weeks. Ten healthy animals were used as controls. At week 7, OA[Osteoarthritis] was surgically induced in left knees of all rabbits. At 22 weeks, after sacrifice, microstructure parameters were assessed by micro-computed tomography, and osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand (RANKL), alkaline phosphatase (ALP) and metalloproteinase 9 (MMP9) protein expressions were evaluated by Western Blot at subchondral bone. In addition, cartilage damage was estimated using the histopathological Mankin score. Significant difference was established at P < 0.05. Subchondral bone area/tissue area, trabecular thickness and polar moment of inertia were diminished in OPOA knees compared with control or OA knees (P < 0.05). A decrease of plate thickness, ALP expression and OPG/RANKL[Osteoperosis and Osteoarthritis alter the OPG/RANKL gradient, and they are also studying subchondral/epiphyseal bone so there is an OPG/RANKL gradient outside the individual osteons] ratio as well as an increased fractal dimension and MMP9 expression occurred at subchondral bone of OA, OP and OPOA knees vs. controls (P < 0.05). In addition, the severity of cartilage damage was increased in OPOA knees vs. controls (P < 0.05). Remarkably, good correlations were observed between structural and remodelling parameters at subchondral bone, and furthermore, between subchondral structural parameters and cartilage Mankin score. Microstructure impairment at subchondral bone associated with an increased remodelling aggravated cartilage damage in OA rabbits with previous OP. Our results suggest that an increased subchondral bone resorption may account for the exacerbation of cartilage damage when early OA and OP coexist simultaneously in same individuals."

So, changes in the OPG/RANKL gradient affect osteoarthritis allowing for bone formation where it didn't occur before.  OPG/RANKL fields occur in the subchondral/epiphyseal bone, the growth plates, bone fractures(microfactures), and in the osteons.  OPG/RANKL fields are modulated by blood cells and hormones(like PTH).

LIPUS and PEMF can also manipulate these fields.  Even though this study involves osteoblasts.  OPG/RANKL magnetic fields are key because they can control bone deposition and could possibly result in new bone being deposited on the longitudinal ends of the bones making you taller!  Endochondral ossification does involve OPG and RANKL signaling but if it were possible to induce OPG/RANKL signaling without a chondrogenic template it might be possible to grow taller in new ways that don't involve a cartilagenous template.

Comparison of low-intensity pulsed ultrasound and pulsed electromagnetic field treatments on OPG and RANKL expression in human osteoblast-like cells.

"LIPUS or PEMF was applied to Saos-2 cells for 10 minutes or 3 hours. RANKL and OPG expressions were analyzed at 0, 4, 8, or 12 hours after treatment with real-time PCR. Secreted protein levels in culture supernatant were analyzed at the same posttreatment time points using specific ELISA assays.
Neither LIPUS nor PEMF had an effect on RANKL protein expression. OPG protein was significantly increased by LIPUS after 0 and 4 hours (brief short-term effect) and was increased almost 2.5-fold by PEMF after 8 hours.[PEMF increases OPG protein more than LIPUS but that is to be expected as OPG involves electromagnetic fields] The mRNA levels of OPG and RANKL were hardly affected by LIPUS treatment at any time point. PEMF induced a fivefold increase in RANKL mRNA expression at t = 0. A brief PEMF treatment of 10 minutes resulted in downregulation of RANKL expression after 0 and 4 hours and upregulation at 12 hours. OPG mRNA was downregulated after 8 hours.
The effects of LIPUS or PEMF expression on OPG and RANKL are limited. From our experiments, it seems that LIPUS treatment resulted in a quick protein response, while the response of cells to PEMF (3 hours) was delayed. The increase in OPG protein at 8 hours post PEMF treatment is indicative of reduction of osteolysis."

PEMF affects the OPG/RANKL gradient more than LIPUS.  If you can use PEMF to induce OPG expression at the longitudinal ends of the bones than you could grow taller.  To do that you'd need osteoblasts at the longitudinal ends of the bones.  So to grow taller with a PEMF/OPG method you have to get the osteoblasts in the right spot and then send the Electromagnetic signals to induce bone formation.

"A pulsed electromagnetic field generator (Physio-Stim, Orthofix Inc, McKinney, Tex) was used for stimulation[Here's a kind of electromagnetic stimulator but not sure if it's sufficient: Platinum Digital TENS Machine and Muscle Stimulator Dual Combo.]. The magnetic field waveform consists of bursts of triangular pulses with a pulse frequency of 3.8 kHz, a burst duration of 5.56 ms, and a burst on-off period of 67 ms. The resulting burst on-off frequency is 15 Hz. The maximum amplitude of the magnetic field was approximately 2 mT (20 G). Electromagnetic field stimulation was applied for 10 minutes or 3 hours. The culture flasks were placed in central position of the generator. After PEMF stimulation, the cells were cultured for 0, 4, 8, or 12 hours. "

<-Now the problem is that this was directly on the osteoblastic cells, we want to induce bone formation in a specific location(on the top of the bone) and we don't know how to do that.  Also, we don't know the other effects of PEMF.

Further, evidence that OPG/RANKL operates based on charge and if we manipulate this gradient so that the osteoblasts are aligned to the subchondral plate we can also grow taller via that mechanism.

Osteoprotegerin is produced when prostaglandin synthesis is inhibited causing osteoclasts to detach from the surface of mouse parietal bone and attach to the endocranial membrane.

"Osteoclast differentiation and activation is controlled, at least in part, by the counterbalancing influences of osteoprotegerin ligand (OPGL) and osteoprotegerin (OPG). We test the hypothesis that OPG mediates the inhibition of osteoclast activity that occurs with indomethacin in the mouse calvaria. Recombinant human OPG, like indomethacin, was found to cause osteoclasts to detach from the bone surface and attach to the adjacent endocranial membrane (periosteum)[OPG can make osteoclasts attach to the periosteum, imagine if there was something that could make osteoblasts attach to the subchondral plate. Recombinant human OPG also inhibited the stimulatory effect of prostaglandin E2 (PGE2)[Remember avocados and green tea can downgrade prostaglandin E2 and possibly other prostaglandins], parathyroid hormone (PTH), and 1,25-dihydroxyvitamin D3 (1,25D3) on osteoclast adhesion to bone after an incubation with indomethacin. A function-blocking antibody to OPG and soluble human OPGL both inhibited the effect of indomethacin, leaving active osteoclasts on the bone[without OPG osteoclasts stayed on the bone itself]. OPG activity was detected in the culture medium from indomethacin-treated bones and PTH, PGE2, 1,25D3, and dexamethasone all inhibited the production of OPG activity. We conclude that, in the absence of specific stimulators of bone resorption, OPG is produced by the mouse calvaria in vitro, which inhibits bone resorption by causing osteoclasts to detach from the bone surface."

OPG made osteoclasts bind to the periosteum rather than the bone.  OPG may just prevent osteoclasts from binding to the bone rather than making the periosteum a more attractive target for binding.

How does RANKL affect osteoblast and osteoclast binding?

Increase in expression of receptor activator of nuclear factor kappaB at sites of bone erosion correlates with progression of inflammation in evolving collagen-induced arthritis.

"The receptor activator of nuclear factor kappaB (RANK)/RANK ligand (RANKL) pathway is critical in osteoclastogenesis and bone resorption and has been implicated in the process of focal bone erosion in arthritis.
DBA-1 mice were immunized with bovine type II collagen/Freund's complete adjuvant and were given an intraperitoneal booster injection of type II collagen on day 21. Arthritis was monitored visually, and joint pathology was examined histologically. RANK and RANKL expression were analyzed using specific immunohistochemistry, and tartrate-resistant acid phosphatase (TRAP) staining was performed. In addition, TRAP and cathepsin K messenger RNA expression were analyzed by in situ hybridization.
A marked increase in the number of cells expressing RANK correlated with the progression of synovial inflammation and clinical disease severity in evolving collagen-induced arthritis (CIA). Interestingly, RANK expression demonstrated a gradient pattern with increased numbers of RANK-positive cells within the synovial infiltrate in areas closer to periosteum and cortical bone[RANK expression is higher in the periosteum]. Cells expressing RANK included cells in synovial tissue[likely the fibrous capsule which is an extension of the periosteum], bone lining cells on the surface of trabecular bone at sites of erosion[bone erosion sends signals for bone modeling to occur], and cells in periosteal areas adjacent to synovial inflammation. In areas where RANK-positive cells were abundant, TRAP-positive, multinucleated osteoclast-like cells were also present at sites of focal bone erosion, suggesting differentiation of synovially derived RANK-positive osteoclast precursor cells into osteoclasts. In addition, TRAP- and cathepsin K-double-positive osteoclast-like cells were detected on the synovial side of cortical bone at sites of early and advanced cortical bone erosion. Sites of RANK expression also correlated well with sites of RANKL expression, and there was a close correlation of the temporal expression of the receptor-ligand pair.
Cells expressing RANK increased in abundance with the progression of arthritis in evolving CIA, and sites of RANK-expressing cells correlated with sites of TRAP-positive, multinucleated osteoclast-like cells as well as with sites of RANKL expression."

Since RANKL expression is higher in the periosteum that could be why osteoclasts become attracted to the periosteum when OPG expression increases.  OPG/RANKL expression may affect osteoclast movement patterns more than osteoblast patterns which will alter the height increase applications of manipulation of this gradient.  You'd have to use osteoclasts to create space in the bone for new cartilagenous templates.


"RANK-positive chondrocytes were detected in articular cartilage of the patella, femur, and tibia in nonarthritic and arthritic mice. However, no RANK expression was found in the growth plate chondrocytes"<-the chondrocytes in the articular cartilage are RANK positive but the chondrocytes in the growth plate are not.  Osteoclasts play an important role during growth by modeling the extracellular matrix.  Articular and growth plate chondrocytes originate from the same cartilagenous template so maybe the change in RANK expression is due to environment.