Gain in Height by Controlling Bone Shape

There's a reason why some bones are shaped differently than others.  Exploiting those differences can be a way to gain in height and grow taller.

Regulation of leg size and shape: Involvement of the Dachsous-fat signaling pathway.

"The protocadherin Dachsous-Fat (Ds-Ft) signaling pathway has been found to be essential for wing development of the fly and leg regeneration of the cricket. The Ds-Ft signaling pathway is linked to the Warts-Hippo (Wts-Hpo) signaling pathway, leading to cell proliferation. Wts-Hpo signaling pathway is involved in the control of organ size, and that this pathway is regulated by Ds-Ft and Merlin-Expanded, which are linked to morphogens such as decapentaplegic/bone morphogenic protein, Wingless/Wnt, and epidermal growth factor[these pathways control anabolic and height growing proteins like WNT and BMPs, it may ultimately be more beneficial to try to manipulate the Ds-Ft pathway rather than all the individual pathways]. steepness of the Ds-Ft gradient controls leg size along the proximodistal axis[So make the Ds-Ft gradiant steeper to grow taller]"

"Hundreds of genetic variants in at least 180 loci influence adult height. The 180 loci are enriched with genes that underlie growth plate development, including hedgehog (Hh) and TGFβ signaling"<-Change TGF-Beta signaling and grow taller(TGF-Beta signaling is enhanced by LSJL)

"Ft is a large atypical cadherin molecule (>5,000 amino acids) as a transmembrane receptor with 34 cadherin domains in its extracellular region"
 
"These phenomena suggest two important points: (1) when cells with non-continuous PVs[Positional Values] are placed next to each other, the missing PVs are intercalated by growth to provide a set of continuous PVs; (2) the gradient in PVs also specifies planar cell polarity (PCP). It is known that intercalary regeneration also occurs in a circumferential direction"<-alter positional values to grow taller

"the direction of slope or vector of the gradient normally determines polarity, then the steepness could measure dimension and regulate growth, referred to as “Steepness model”. An important aspect of this model is that dimension sensing depends on the linear gradient of PVs established between the boundaries of a defined and growing population of cells whose maximum and minimum scalar values are constant; consequently, as the organ grows, the gradient becomes less steep"<-increase the steepness of the gradient to grow taller.  To do that increase specific positional values.  LSJL may do this to some degree by only loading the epiphysis and thus increasing the positional values relative to the rest of the bone.

So increasing the positional values is a way to grow taller and if specific positional values are increased the rest of the positional values will be intercalated by the body.

Methodology of increasing positional values is unclear with most of the research being done on the Hydra.  It's possible however that LSJL alters the positional value of bone cells.

A good way to study positional value would be muscular hypertrophy.  Are there any individual parts of the muscle that grow disproportionately?  Bodybuilders often go for the peak but I haven't seen any specific examples of a bodybuilder who developed a bicep peak.  If we know what stimulus they used to induce specific growth in only parts of the muscle we can extrapolate that to other tissue types(bone, cartilage).

Here's a paper explaining how chondrocyte cell polarity may control height growth:

Cell polarity: The missing link in skeletal morphogenesis?

"the growth rates and mechanical properties of each type of element are distinct, even though the development of each [skeletal] element is regulated by a common signaling network "<-So how does endochondral ossification manage to produce so many different bone shapes?

"each limb develops independently from both the contra-lateral limb and the rest of the body, yet during outgrowth the corresponding limbs are nearly identical in size."<-so there must be some factors regulating the growth of both limbs.

"At [the] early stages of skeleton formation, the morphology of the cartilage grossly resembles that of the final bony element."<-So it could be the initial shape of the cartilage that determines the final growth rather than chemical signaling.

"In the growth plate, chondrocyte maturation occurs non-uniformly and with a distinct polarity within the cartilage element, such that cells closer to the center of the [skeletal] element are more mature than chondrocytes nearer the joint surface"<-can this polarity regulate height growth?

"During limb bud formation and elongation, tissue polarity is established along three axes (anterior-posterior, proximal-distal and dorsal-ventral) by the interactions of several signaling pathways. The proximodistal axis is initially established when Hox genes initiate expression of fibroblast growth factor (FGF) and Wingless/int-1 (Wnt) genes (that encode secreted signaling molecules) in the limb field mesenchyme, which, in turn, activates FGF expression in the overlying ectoderm."<-the proximaldistal axis would be the one that controls height.

"Interactions between the mesoderm and ectoderm of the limb bud induce formation of the apical ectodermal ridge (AER), an epithelial structure that promotes limb bud outgrowth and sequentially expands the cell populations that form distinct segments of the limb skeleton"<-Can we mimic the apical ectodermal ridge to induce post-pubertal height growth?

"Distal FGF signaling is opposed by proximal retinoic acid, a small molecule produced by the flank mesenchyme. Retinoic acid diffuses into the limb bud and is degraded by distal mesenchyme cells, thus creating a proximal-to-distal morphogen gradient[Hyaluronic Acid has been shown to significantly increase MSC expression of retinoic acid]. The importance of these morphogen gradients to limb patterning was recently underscored by in vitro studies demonstrating that proximodistal identity is expressed when limb mesenchyme cells experience retinoic acid signaling after leaving the distal FGF signaling domain."<-So since hyaluronic acid can increase levels of retinoic acid and retinoic acid can affect limb patterning then hyaluronic acid supplementation can affect limb patterning.

"Limb outgrowth is driven by cell proliferation that requires FGF and Wnt5a signaling in the distal mesenchyme."

"SHH[Sonic Hedgehog] secretion by the ZPA[Zone of polarizing activity] also forms a concentration gradient across the limb bud, which patterns the mesenchyme along the anteroposterior axis."<-so concentration gradients affect mesenchymal patterning thus things that affect hydrostatic pressure like LSJL might also be able to affect the patterning of mesenchyme.

"observations suggest that regulation of cell shape, oriented cell division and directional cell movement might promote growth and morphogenesis in the limb bud."<-so we can manipulate cell shape(done by hydrostatic pressure), cell division(done by growth factors), and directional cell movement(done by disrupting the actin cytoskeleton) to manipulate growth.

"columns form in a two-step process from cells in the resting zone. In the first step, elliptical resting chondrocytes without oriented cell behaviors progressively flatten and display planar alignment of the mitotic spindle and the cell axes[We can flatten cells with various stimuli].  In the second phase, daughter cells that were displaced laterally by cell division reorient 90° to form a column parallel to the long axis of the skeletal element[this displacement may upregulate genes that cause the formation of the growth plate]. Interestingly, the organization of proliferative chondrocytes is also evident at the subcellular level in the planar alignment of the primary cilium of each chondrocyte at the center of the column[cilium are sensory organelles thus sensory functions are important to growth, which indicates that chondrocyte organization is important to growth].  Thus, as in the limb bud, cartilage growth is associated with planar alignment of cell axes, oriented cell division and oriented cell movements."

"Chondrocytes in the resting zone are elliptical and do not exhibit a bias in the orientation of cell division. After being recruited into the proliferative zone, however, chondrocytes demonstrate planes of cell division that are perpendicular to the long axis of the skeletal element. The resultant daughter cells then rearrange to form a clonal column parallel to the long axis of the growth plate."<-so the reason why long bones become long and not wide is an orientational bias of cell division.

"in the absence ofWnt5a function, limb mesenchyme cells adopted an elliptical shape, and the long axis of the cell was not primarily oriented toward the ectoderm."<-So the Wnt5a gene is primarily responsible for determining the orientational bias of cell division.

"Depending on the cellular context, non-canonical Wnt signaling can function by regulating intracellular Ca2+and/or activating proteins such as the GTPase Rac and the intracellular kinases Src, calcium/calmodulin-depenent kinase II (CAMKII) or Jun N-terminal kinase (JNK). These pathways regulate diverse cellular processes, including cell polarity, dynamic organization of the cytoskeleton and gene expression."<-We can manipulate these pathways directly to alter height growth.

"experimental manipulations that alter cell shape, cell arrangement or oriented cell movement also alter growth vectors and result in limb structures that are shorter and wider (thicker) than in wild type."<-so we alter cell shape, arrangement, and movement to build limb structures that are longer.

"hypertrophy generates the driving force for bone elongation."

"column formation in discoid proliferative chondrocytes [may serve] to maximize cell density in the longitudinal axis while regulating cell density (i.e., column number) in the lateral axis. The net effect of this arrangement is to constrain the growth-promoting effects of hypertrophy to the longitudinal axis. Similarly, although chondrocytes of the elongating digits do not appear to form columns and do not undergo hypertrophy in the early stages, cell polarity might still play a role in digit lengthening. The maintenance of cell shape and orientation of cell division could generate a driving force for digit elongation, even if together these cell behaviors simply limit lateral expansion of the developing digit cartilage."<-so cell proliferation merely causes hypertrophic chondrocytes to generate more length rather than width?  If cell proliferation was reduced then the bones would be the same size but would be wider rather than longer.  This could explain an ectomorphic versus a mesomorphic body type.  Ectomorphs have a lot cell proliferation and a high level of hypertrophic chondrocytes whereas mesomorphs have a high level of hypertrophic chondrocytes but relatively low levels of cellular proliferation.

So to manipulate height growth we alter cell shape, orientiation, and division.  We can alter cell shape and division of the mesenchyme post puberty and thus grow taller after puberty.  During puberty, if you want to be tall and thin focus more on increasing chondrocyte cell proliferation.  Whereas, if you want to be broader then focus more on increasing chondrocyte hypertrophy.

Here's another article about Wnt5a and it's importance in growth plates.  This article was co-authored by R. Tracey Ballock.

Activation of Wnt planar cell polarity (PCP) signaling promotes growth plate column formation in vitro.

"Disrupting the Wnt Planar Cell Polarity (PCP) signaling pathway in vivo results in loss of columnar growth plate architecture. We hypothesized that activation of the Wnt PCP pathway ingrowth plate chondrocyte cell pellets would promote columnar organization in these cells that are normally oriented randomly in culture. Rat growth plate chondrocytes were transfected with plasmids encoding the Fzd7 cell-surface Wnt receptor, a Fzd7 deletion mutant lacking the Wnt-binding domain, or Wnt receptor-associated proteins Ror2 or Vangl2, and then cultured as three-dimensional cell pellets in the presence of recombinant Wnt5a or Wnt5b for 21 days. Cellular morphology was evaluated using histomorphometric measurements. Activation of Wnt PCP signaling components promoted the initiation of columnar morphogenesis in the chondrocyte pellet culture model, as measured by histomorphometric analysis of the column index (ANOVA p = 0.01). Activation of noncanonical Wnt signaling through overexpression of both the cell-surface Wnt receptor Fzd7 and receptor-associated protein Ror2 with addition of recombinant Wnt5a promotes the initiation of columnar architecture of growth plate chondrocytes in vitro, representing an important step toward growth plate regeneration. "

So Wnt5a, Ror2, and Fzd7 are all important to heigth growth.

"Grafting of chondrocyte-seeded scaffolds into proximal tibial growth plate defects led to formation of isolated pockets of chondrocyte columns by day 14 in rabbits"

"the cellular implants failed to completely restore the columnar architecture and function of a growth plate in vivo, however, which is a prerequisite for maintenance of normal long-term growth."<-Likely due to no Wnt5a.

"Wnt signaling consists of two major arms, the canonical and noncanonical pathways. While the canonical Wnt pathway operates through cytoplasmic accumulation of β-catenin in order to regulate gene transcription, the noncanonical pathways transduce their signals in a β-catenin independent fashion. The noncanonical Wnt pathways include the Wnt PCP and Wnt Ca2+ pathways. Although many of the established Wnt ligands and receptors have been found to participate in multiple Wnt signaling pathways, the downstream effects of the canonical and noncanonical arms of Wnt signaling are markedly different. In chondrocytes, canonical Wnt signaling inhibits chondrogenesis and promotes hypertrophy, while the noncanonical Wnt PCP pathway is thought to influence cellular morphology[so want the noncanonical non-Beta-catenin Wnt signaling]. The Wnt Ca2+ pathway has been implicated in the regulation of chondrocyte maturation and proliferation through direct and indirect inhibition of canonical Wnt signaling, but is not the focus of the present study. Wnt PCP effector pathways alter the actin cytoskeleton and cellular morphology through downstream activation of the small GTPases Rac and Rho, and establish cell polarity in response to concentration gradients of appropriate Wnt ligands."

"Wnt PCP components are necessary for flattened chondrocyte morphology and cell stacking in the developing chick limb, and for proper limb elongation in mice. In chicks, maintenance of columnar architecture and flattened cell morphology of proliferative zone chondrocytes is dependent upon expression levels of the Wnt receptor-associated proteins Fzd7 and Vangl2."<-Thus to restore growth plates we have to restore Wnt PCP components.

"Treatment of chondrocyte pellet cultures with Wnt5a or Wnt5b added to the culture media promoted cell flattening, resembling the appearance of proliferative zone chondrocytes."<-So after forming growth plates with LSJL.  We have to upregulate Wnt5a.  Too bad we don't have data on whether LSJL is inducing chondrogenesis but not growth plates.

"Wnt5a and Fzd7 had a marked impact on induction of columnar architecture, and regions of these pellets displayed architecture reminiscent of the native growth plate"<-So upregulating Fzd7 would be nice to regenerate growth plates as well.

"Fzd7 overexpression and Wnt5a, Wnt5b treatments promote cell stacking."

"Wnt5a promotes the formation of a trimeric receptor complex composed of Fzd7, Ror2, and Vangl2"<-So manipulating any of those 4 likely helps height growth

"Vangl2 misexpression (both under- and overexpression) disrupts column formation in vivo."

According to Transcriptional mechanisms of WNT5A based on NF-κB, Hedgehog, TGFβ, and Notch signaling cascades, TGF-Beta signals upregulate Wnt5a.  And since LSJL upregulates TGF-Beta it may already upregulate Wnt5a.

ROLES OF THE FAT PATHWAY IN CARTILAGE PATTERNING AND POLARITY

"Our recent finding that Rere, a close relative of the transcription factor Atrophin which regulates Dachsous (Dchs)-Fat signaling in flies, is required for craniofacial development in zebrafish has implicated the Fat pathway in planar cell polarity (PCP) during skeletogenesis. Dramatic results from our laboratory now demonstrate an even more profound role for a Fat3 in cartilage differentiation, consistent with PCP. Embryos deficient in Rere or Fat3 develop cartilages in which stacking and differentiation are uncoupled in the pharyngeal arches and this leads to joint fusion. Moreover, Rere and Fat3 are critical for coordinating responses of skeletal progenitors to Bmp and Fgf signaling during craniofacial development. We propose to explore the roles of the Fat pathway and PCP in cartilage and joint formation in three sets of experiments. The first aim will address the hypothesis that the Fat pathway couples cartilage stacking and differentiation. Cartilage and joint phenotypes will be evaluated in Rere and Fat3 mutants using live imaging. We will identify the Dchs ligand(s) for Fat in cartilage and create ectopic sources to study signal propagation and modulation{an ectopic source could possibly be a new growth plate}. We will also compare the activities of the Fat pathway and Wnt/PCP and their interdependence in cartilage development. The second aim will address the hypothesis that the Fat pathway coordinates polarized responses to Bmps and other growth factors, thereby linking early patterning events with later organ formation. For this we have new markers of cytoskeletal polarity and cilia that reveal unexpected boundaries of polarity in skeletal progenitors. Finally, the third aim will focus on joint formation and test the hypothesis that the Fat pathway prevents cartilage stacking and differentiation in the joint interzone with inducible elimination or overexpression of Rere or Fat3."