EPH receptors

LSJL downregulates EphrinB3.  It upregulates Epha3 and Epha5

EphB/ephrin-B interactions mediate human MSC attachment, migration and osteochondral differentiation.

"EphB and ephrin-B molecules were expressed by ex vivo expanded human MSC populations and within human bone marrow trephine samples. To elucidate the contribution of EphB/ephrin-B molecules in MSC recruitment, we performed functional spreading and migration assays and showed that reverse ephrin-B signalling inhibited MSC attachment and spreading by activating Src-, PI3Kinase- and JNK-dependent signalling pathways. Forward EphB2 signalling promoted MSC migration by activating the Src kinase- and Abl-dependent signalling pathways.  Activation of ephrin-B1 and/or ephrin-B2 molecules expressed by MSC was found to increase osteogenic differentiation, while ephrin-B1 activation promoted chondrogenic differentiation. These observations suggest that EphB/ephrin-B interactions may mediate the recruitment, migration and differentiation of MSC during bone repair."

"EphB2, EphB4 and ephrin-B1 expressions appeared most pronounced within the human bone, localising to bone lining cells, "

"ephrin-B3 was expressed by a large number of bone marrow cells"<-the downregulation of ephrinB3 in LSJL could be a sign of bone marrow cell differentiation.

"Blocking the EphB2 receptor, significantly inhibited GAG synthesis"

"the formation of a PDZ complex containing NHERF1, PTPN13 and TAZ with ephrin-B1 results in the dephosphorylation of TAZ, which allows its translocation into the nucleus and subsequent activation of osteogenic specific transcription factor, Osterix"

Loss of Eph-receptor expression correlates with loss of cell adhesion and chondrogenic capacity in Hoxa13 mutant limbs.

"In mice that lack the Hoxa13 gene, the mesenchymal condensations that form the autopod skeletal elements are poorly resolved, resulting in missing digit, carpal and tarsal elements. In addition, mesenchymal and endothelial cell layers of the umbilical arteries (UAs) are disorganized, resulting in their stenosis and in embryonic death.  We generated a loss-of-function allele in which the GFP gene was targeted into the Hoxa13 locus. This allele allowed FACS isolation of mesenchymal cells from Hoxa13 heterozygous and mutant homozygous limb buds. Hoxa13(GFP) expressing mesenchymal cells from Hoxa13 mutant homozygous embryos are defective in forming chondrogenic condensations in vitro. Analysis of pro-adhesion molecules in the autopod of Hoxa13 mutants revealed a marked reduction in EphA7 expression in affected digits, as well as in micromass cell cultures prepared from mutant mesenchymal cells. Finally, antibody blocking of the EphA7 extracellular domain severely inhibits the capacity of Hoxa13(GFP) heterozygous cells to condense and form chondrogenic nodules in vitro, which is consistent with the hypothesis that reduction in EphA7 expression affects the capacity of Hoxa13(-/-) mesenchymal cells to form chondrogenic condensations in vivo and in vitro. EphA7 and EphA4 expression were also decreased in the mesenchymal and endothelial cells that form the umbilical arteries in Hoxa13 mutant homozygous embryos."

"the addition of the anti-EphA7 antibody dramatically reduced both cell adhesion and chondrogenic nodule formation of Hoxa13GFP heterozygous cells."

MicroRNA-34a regulates migration of chondroblast and IL-1β-induced degeneration of chondrocytes by targeting EphA5.

"We investigated the role of miRNA-34 during chondrogenic differentiation of chick limb mesenchymal cells. Expression of miR-34a increased upon chondrogenic inhibition. Blockade of miR-34a via PNA-based antisense oligonucleotides (ASOs) recovered the chondro-inhibitory actions of JNK inhibitor on migration of chondrogenic progenitors and the formation of precartilage condensation. EphA5 is a relevant target of miR-34a during chondrogenesis. MiR-34a was necessary and sufficient to down-regulate EphA5 expression, and up-modulation of EphA5 is sufficient to overcome inhibitory actions of miR-34 inhibition on cell migration and condensation of chick limb mesenchymal cells on collagen substrate. miR-34a is a negative modulator of chondrogenesis, particularly in migration of chondroblasts, by targeting EphA5 and resulting inhibition of cellular condensation during chondrogenesis of chick limb mesenchymal cells."

"In articular chondrocytes, Wnt-3a caused dedifferentiation of chondrocytes by upregulating the expression of c-Jun and its phosphorylation by JNK, resulting in activation of the c-Jun/activator protein. Treatment with transforming growth factor-β superfamily members promoted cartilage-specific gene expression during in vitro chondrogenic differentiation of mesenchymal progenitor cells from bone marrow and trabecular bone through activation of the mitogen-activated protein (MAP) kinases, p38, ERK-1, and JNK"

"miR-34a is one of key modulators in JNK-induced chondrogenic differentiation"

"EphA5 increases spreading and migration of chondroprogenitors in collagen substrate"

"the protein level of type II collagen decreased when cells were exposed to JNK inhibitors"

"miR-34a was significantly up-regulated by IL-1β and silencing of miR-34a significantly prevented IL-1β-induced down-regulation of type II collagen in rat chondrocytes. Increased expression level were observed in human articular chondrocytes exposed to IL-1β and human chondrocytes were prepared from osteoarthritic zone suggesting the involvement of miR-34a in the dedifferentiation of human articular chondrocytes by IL-1β. Co-treatment of miR-34a inhibitor induced the recovery of EphA5 level and type II collagen protein level which were suppressed by treatment of IL-1β. The expression level of EphA5 and type II collagen was significantly decreased in human chondrocytes were prepared from osteoarthritic zone"

Where the Load is

In a previous entry, I talked about the reasoning which states that if weight lifting has the ability to produce height increase(via bone size increase) then why aren't bodybuilders gaining height? I reasoned that weightlifters(powerlifters, bodybuilders, strongman, and misc) are in fact gaining height but that it is not large enough to be unable to be dismissed by measurement error. Look at how extreme a bodybuilders shoulder and bicep development is relative to how it is compared to non-specific weight training. A normal weight lifter will not have a bodybuilders physique and proportions. Bodybuilders do highly specialized routines to get their disproportionately large arms and shoulders.

So too must people who want to train for height increase via bone increase. Imagine the standard back squat movement. At the top of the movement, the bar is putting most of the load on the bone. In the middle of the movement, most of the load is on the muscle. At slightly below parallel most of the load is on the tendons and ligaments. Same with bench press: Arms locked or near locked out bone, in between muscle, at the chest tendons and ligaments. If you are very flexible, you might have to do dumbell bench to get the load onto the tendons and ligaments.

How to build bone? Perform exercises where most of the load is on the bone(some load will always be on the muscle). Bodybuilders stress how important ROM after all. Bodybuilders want to keep their ROM in the muscle building range, we height builders want to keep our ROM in the bone building range.

PKR

EIF2a-p is important for PKR inhibition of Cyclin D1.

Double-stranded RNA-dependent protein kinase regulates insulin-stimulated chondrogenesis in mouse clonal chondrogenic cells, ATDC-5.

"Double-stranded RNA-dependent protein kinase (PKR) is an interferon-induced protein that has been identified and characterized as a translational inhibitor in an interferon-regulated antiviral pathway. PKR inactivation suppresses osteoblast calcification and osteoclast formation. PKR is required for the in vitro differentiation of the mouse clonal chondrogenic cell line ATDC-5. ATDC-5 cells treated with insulin differentiated into chondrocytes and produced an alcian-blue-positive cartilage matrix. The protein expression of signal transducers and activators of transcription (STAT) peaked at day 7 of differentiation, whereas the expression of SRY-box-containing gene 9 (Sox-9), which is a transcription factor for chondrocyte differentiation, increased gradually. When the cells were treated with a PKR inhibitor (2-aminopurine), the cartilage matrix formation decreased. The protein expression of STAT1{Stat1 interacts with Cyclin D1} continued to increase up to day 21, whereas the expression of Sox-9 was low and did not increase. PKR was localized to a marginal region of the mandibular condyle cartilage in mouse embryos. PKR has important functions in the differentiation of chondrocytes through the modulation of STAT1 and Sox-9 expression."

Since PKR is important to chondroinduction maybe EIF2a-p and therefore Salubrinal are too.

"PKR/eIF-2α cascade has been implicated as a general transducer of apoptosis"

"2-Aminopurine (2-AP), which interacts with the ATP-binding site of PKR, is a potent inhibitor of PKR  and blocks tumor necrosis factor-α (TNF-α) and C2-ceramide [activities]"

"ATDC-5 cells show characteristics of undifferentiated prechondrogenic cells in the growth phase and maintain a potential for chondrogenesis. When treated with insulin, ATDC-5 cells undergo cellular condensation in the post-confluent phase and acquire the phenotype of chondrocytes, i.e., proteoglycan synthesis and type II collagen expression"

"the level of STAT1 protein was higher in the cells treated with insulin, especially at day 7 of culture and inhibition of PKR resulted in the sustained expression of STAT1. PKR interacts with STAT1 and PKR-STAT1 complexes interfere with the transactivation capacity"

"PKR inhibition sustains STAT expression, which results in the inhibition of chondrogenesis."

Growth Taller via Autophagy?

Autophagy may be a way to resist aging related degenerations.  Autophagy, unlike apoptosis, allows for recycling of cellular components so if all the hypertrophic cells in the growth plate underwent autophagy over apoptosis it may be possible to grow taller indefinitely.


Regulation of autophagy in human and murine cartilage: hypoxia-inducible factor 2 suppresses chondrocyte autophagy.

"HIF-2 was expressed abundantly by cells in human and murine articular cartilage and in the cartilage of mineralizing vertebrae from neonatal mice. Protein levels were reduced in articular cartilage from older mice, in end-plate cartilage from mice, and in chondrocytes from human osteoarthritic (OA) cartilage. HIF-2 was robustly expressed in the prehypertrophic cells of mouse growth cartilage. When HIF-2alpha was silenced, the generation of reactive oxygen species was found to be elevated, with a concomitant decrease in catalase and superoxide dismutase activity. Suppression of HIF-2 was associated with decreased Akt-1{pAkt was decreased as well} and mammalian target of rapamycin activities, reduced Bcl-xL expression, and a robust autophagic response, even under nutrient-replete conditions. In these silenced chondrocytes, HIF-1 expression was elevated. Decreased HIF-2 expression was associated with autophagy in OA tissues and aging cartilage samples. The autophagic response of chondrocytes in HIF-2alpha-knockout mouse growth plate showed an elevated autophagic response throughout the plate.

HIF-2 is a potent regulator of autophagy in maturing chondrocytes. HIF-2 acts as a brake on the autophagy-accelerator function of HIF-1."

"The maturing cells in [growth plate] cartilage undergo a series of phenotypic changes which include secretion of a unique sets of proteins into the avascular extracellular matrix, up-regulation of alkaline phosphatase and the release and subsequent mineralization of matrix vesicles. Prior to deletion from the plate, the mature hypertrophic chondrocyte becomes glycolytic and undergoes functional and immunohistochemical changes that are characteristic of autophagy. HIF-1, a transcription factor that responds to the tissue oxemic state, promotes chondrocyte autophagy"

"upregulation of HIF-2 lowers intracellular ROS levels by promoting the activities of the dismutating proteins, catalase and superoxide dismutase. The observation that HIF-2 knockout animals are small suggests that there may be an increased rate of chondrocyte apoptosis that serves to impede normal long bone growth."<-so increased autophagy may not increase height however this may be due to other effects like reduced Akt and MTor.

"When we compared neonatal cartilage in the mineralizing vertebrae with end plate cartilage of older mice a dramatic decrease in HIF-2α expression is seen."

HIF-2 reduces reactive oxygen species.  It's possible that autophagy results in the recycling of everything including things that would reduce height growth.

Oxygen may reduce HIF-2 levels providing the signal for epiphyseal mineralization.

Chondrocytes embedded in the epiphyseal growth plates of long bones undergo autophagy prior to the induction of osteogenesis.

"TOR plays a critical role in autophagy serving as the sensor that integrates metabolic and growth factor signals.  Terminal chondrocytes exhibit autophagic characteristics. Morphologically, the arrested cells contain double membrane vacuoles; there is a loss of membrane structure, limited staining and organelle destruction. Since the life history of the growth plate chondrocyte is very short, even minor disturbances in the metabolic state can result in gross impairment of growth. The induction of the autophagic response, permits the terminally differentiated cells to survive the brief rigors of the harsh local microenvironment."

"in the maturing zone of the growth plate, the central core of cells are most removed from oxygen and may be depleted of growth factors and critical nutrients.  In this hypoxic microenvironment, gradients in nutrients and specific growth factors would be countered to some extent by upregulation of ATP generation through the glycolytic pathway.  The loss of growth factor signaling and nutrient stress would suppress mTOR

and activate autophagy."

The life history of the growth plate chondrocyte is 1-2 days.

Chondrocyte autophagy is stimulated by HIF-1 dependent AMPK activation and mTOR suppression.

"Chondrocytes [express] the energy sensor AMPK-1 and [activation increases] with maturation. Thapsigargin treatment activated AMPK and autophagy in a HIF-1-dependent manner. Using serum-starved AMPK-silenced cells, we demonstrated that AMPK was required for the induction of the autophagic response. We also noted a change in chondrocyte sensitivity to apoptogens, due to activation of caspase-8 and cleavage and activation of the pro-apoptotic protein, BID. To test the hypothesis that AMPK signaling directly promoted autophagy, we inhibited AMPK activity in mTOR silenced cells and showed that while mTOR suppression induced autophagy, AMPK inhibition did not block this activity. Because of the micro-environmental changes experienced by the chondrocyte, autophagy is activated by AMPK in a HIF-1-dependent manner."

"AMPK activity is responsive to hypoxia and hyperosmotic stress. The kinase is sensitive to the AMP/ATP ratio: binding of AMP activates AMPKand induces phosphorylation by the tumor suppressor LKB1. When bound, AMP inhibits it’s dephosphorylation by Protein Phosphatase 2C. AMPK functions as an energy sensor that triggers catabolic pathways that produce ATP, while inhibiting energy-consuming anabolic activities."

"autophagy is stimulated by HIF-1 and that in concert with chondrocyte maturation, there is a profound decrease in ATP and a concomitant rise in AMP, key regulators of AMPK"

Mice growth plate cartilage cells were used.

"epiphyseal chondrocytes express AMPKa1".  AMPKa2 is expressed by osteoblasts but not chondrocytes.

"Thapsigargin is an inhibitor of sarco/endoplasmic reticulum Ca2+ ATPases (SERCA). It raises the cytosolic calcium concentration by blocking the ability of the cell to pump calcium into the sarcoplasmic and endoplasmic reticula (ER; as a result, these stores become depleted, resulting in ER stress and ultimately cell death. In a number of tissues, AMPK is activated both by a decrease in energy charge and by an increase in intracellular calcium flux."

"AMPK regulates the induction of autophagy by modulating the association between Bcl-2 and Beclin-1"

"AMPK suppression leads to a significant activation of Akt-1."

"While AMPKa1 positive cells were seen throughout the growth plate, the most densely stained cells were localized to the pre-hypertrophic and hypertrophic regions of the epiphysis. Remarkably, the location of the maximally positive chondrocytes was similar to that described for both Beclin-1 and LC-3, two major indicators of autophagic vacuole formation"

" when HIF-1 was silenced, AMPK activation was blocked; in contrast, there robust AMPK activation when control chondrocytes were treated with thapsigargin."

"autophagy can serve to delay the onset of apoptosis"

How does Salubrinal affect autophagy?

Endoplasmic reticulum stress triggers autophagy in malignant glioma cells undergoing cyclosporine A-induced cell death.

"Autophagy is a conserved, self-digestion process that is activated in response to nutrient limitation but acting also as an alternative death mechanism under certain conditions. It is accompanied by the progressive formation of vesicle structures from autophagosomes to autophagolysosomes orchestrated by autophagy effectors (Atg proteins) and modulators (that is, mTOR-mammalian target of rapamycin as a negative regulator). We demonstrate that cyclosporine A (CsA, an immunophilin/calcineurin inhibitor) induces cell death with some apoptotic features but also accompanied by the appearance of numerous cytoplasmic vacuoles, immunostained for endoplasmic reticulum (ER) and autophagy markers. The induction of ER stress in glioma cells by CsA was evidenced by detection of unfolded protein response activation (phosphorylation of PERK, accumulation of IRE1α) and accumulation of ER stress-associated proteins (BIP and CHOP). Formation of the acidic vesicular organelles, increase of autophagic vacuoles, GFP-LC3 punctation (microtubule-associated protein light chain 3) and LC3-II accumulation upon CsA treatment confirmed activation of autophagy. Decrease of phosphorylation of 4E-BP1, p70S6K1 and its downstream target S6 ribosomal protein demonstrate inhibition of mTOR signaling by CsA. Salubrinal and silencing of PERK and IRE1α partially blocked CsA-induced accumulation of LC3-II{So Salubrinal may inhibit autophagy}. It suggests that ER stress precedes CsA-induced autophagy. Surprisingly, silencing of autophagy effectors ULK1, Atg5 or Atg7 increased the level of active caspases 3, 7 and PARP degradation in CsA-treated cells. CsA induces both apoptosis and autophagy in malignant glioma cells via induction of ER stress and inhibition of mTOR/p70S6K1 pathway, however autophagy is cytoprotective in this context."

"In mammalian cells, PERK (protein kinase-like endoplasmic reticulum kinase), IRE1 (inositol-requiring enzyme 1) and ATF6 (activating transcription factor 6) sense accumulation of unfolded proteins in the ER lumen and activate specific cellular response. PERK phosphorylates the α subunit of the translation initiation factor, eIF2α, which attenuates general protein synthesis and initiates the selective translation of some stress-responsive transcripts. IRE1 is a serine/threonine kinase and also an endoribonuclease, which catalyzes the splicing of the mRNA encoding the transcription factor XBP-1 (X-box binding protein 1). PERK mediates transcriptional activation of LC3 and Atg5 proteins under hypoxic conditions via the transcription factors ATF4 (activating transcription factor 4) and CHOP (C/EBP homologous protein), respectively."

According to Effects of salubrinal on cadmium-induced apoptosis in HK-2 human renal proximal tubular cells., Salubrinal protected from apoptosis but not autophagy.  This may be due to the fact that in this study Salubrinal did not increase ATF4 levels.

ASK1 overexpression accelerates paraquat-induced autophagy via endoplasmic reticulum stress.

"Apoptosis signal-regulating kinase 1 (ASK1) is activated by various types of stress, including, endoplasmic reticulum (ER) stress. ASK1 executes apoptosis and/or autophagy under ER stress. [We used] SH-SY5Y cells overexpressing wild-type (WT) ASK1. We show an important autophagic response and an acceleration of the paraquat (PQ)-induced autophagy with hallmarks as accumulation of autophagic vacuoles, activation of beclin-1, accumulation of LC3 II, p62 degradation, and mammalian target of rapamycin dephosphorylation. Inhibition of autophagy caused an exacerbation of the apoptosis induced by WT ASK1 overexpression{So the question is which would increase height more autophagy or apoptosis?} with or without PQ.  Autophagic response could have a protector role. We found an increase in the phosphorylation of the proteins such as IRE1 and eIF2α in response to both the overexpression of WT ASK1 and pesticide exposure.WT ASK1 overexpression-induced autophagy is an event that occurs in parallel with ER stress activation. The importance of ER stress in the autophagy induced by ASK1 and/or PQ was confirmed with salubrinal, a selective inhibitor of eIF2α dephosphorylation. PQ induces an early ER stress response that is correlated with the activation of autophagy as a protective response, which is accelerated in cells that overexpress WT ASK1."

"Apoptosis signal-regulating kinase 1 (ASK1) is a member of the mitogen-activated protein kinase family (MAP3K) that activates the c-jun N-terminal kinase (JNK) and p38 pathways by directly phosphorylating, and thereby activating, their respective MAP2Ks (also called mitogen activated kinase kinase [MKK]), MKK4(SEK1)/MKK7, and MKK3/MKK6"

"preincubation with [salubrinal] rescued cells from the apoptotic event (release of cytochrome c from mitochondria into the cytosol, caspase-3 activation, and nuclear fragmentation) induced by exposure to PQ, the overexpression of WT ASK1, and the combination of both"

Misc Stem Cell types to chondrocytes

Chondrogenic Differentiation of Amniotic Fluid Stem Cells and Their Potential for Regenerative Therapy.

"Chondrocytes constituting articular cartilage show some functional and structural differences from growth plate chondrocytes. Whereas the latter undergo highly dynamic changes as long bone growth persists, articular cartilage chondocytes provide continuous function throughout life. Articular chondrocytes originate from a pool of growth differentiation factor 5 (GDF5) positive progenitor cells and that growth plate chondrocytes are derived from GDF5 negative cell populations"

"GDF5 expression is first evident in the mesenchymal interzone of early joints. The interzone is an essential regulator region and it has been widely assumed that it is responsible for formation of joint structures including articular cartilage, ligaments and synovial lining. Interzone cells express a number of genes able to control joint development, like: GDF5, WNT5A, WNT4, GLI3{up in LSJL}, CD44, ERG{up} and NOGGIN. GDF5 has chondrogenic activity, whereas WNT ligands and NOGGIN are anti-chondrogenic"

"Joint cavitation involves differential hyaluronan synthesis under the influence of mechanical stimuli. The hyaluronan receptor, CD44, is expressed at the joint interzone and at developing articular surfaces and it induces cell adhesion as well as cell separation, depending on the concentration of hyaluronan"

"Mature articular cartilage consists of three zones: the superficial zone, where small and flat chondrocytes reside and collagen fibrils are orientated parallel to the surface, the transitional zone, representing the thickest part of cartilage, where collagen fibers are less organized, and the radial zone, where fibers are orientated perpendicular to the surface and oxygen tension is below 1 %. Underneath is the tidemark, which delineates hyaline articular cartilage from calcified cartilage."

" Upon hypertrophy, growth plate chondrocytes downregulate SOX9 and type II collagen expression, whereas RUNX2, type X collagen, alkaline phosphatase and VEGFA are upregulated"

"[Amniotic Fluid Stem Cells] can be expanded in cell culture and show expression of CD29, CD44, CD73, CD90, CD105 and SSEA4 with over 90 % of cells being positive for OCT4"

"supplementation of chondrogenesis promoting factors is necessary for successful manipulation of AFS cells and that the isolation of c-kit positive cells also enhances the differentiation process."

"GDF5 over-expression results in cartilage overgrowth and joint fusion in vivo"

"WNT-4, WNT-14, WNT-16 and the WNT signaling mediator β-catenin also show anti-chondrogenic activity during joint formation"

"up regulation of HIF1α (hypoxia-inducible factor) and HIF2α is essential for SOX9 expression"

"In articular cartilage autophagy plays an important role, since it protects chondrocytes from age related cell death and preserves homeostasis within the tissue{perhaps in growth plate chondrochytes as well}. During aging-related intracellular changes, biochemical stimuli as well as mechanical stress can lead to OA and during this process the competence to undergo autophagy is lost"<-mTor is involved in the regulation of growth plate autophagy.  Proteoglycan sulfation can determine the efficience of chondrocyte autophagy.

"The induction of autophagy is able to prevent ageing related degenerations."<-maybe autophagy can prevent epiphyseal fusion since epiphyseal fusion is an aging related degeneration.

Analysis of Common Height Increase Methods

Hypnosis-You have to denature the bone in order to gain height. Hypnosis doesn't do that.  It is possible that if hypnosis increases the amount of growth hormone in your body for your to grow taller via greater periosteal width of short and irregular bones.  Also, if you are hypnotized while laying on your back you can gain temporary height by temporarily decompressing the spine(laying on your back results in the height gain not the hypnosis).

Stretching-Stretching usually affects the muscle not the bone(perhaps the bone in a minor way). You can have the longest muscles in the world but they will look stocky because they are resting on short bones.  You need to apply stretching forces on your bones for example by putting weights on the ends of the bones and longitudinally stretching them.

MMA, Boxing, Football-There have been reports of people having fractures of bones and denaturing the bone during the healing and thus being taller. However, their are risks in these sports, it's the physical act of being tackled in football that causes the fractures needed to denature the bone and there is no guarantee that the damage to yourself will not be too severe.

As for boxing/kickboxing, the opponents are not aiming for your bones. They are aiming for your liver or your brain.  Heavy bag punching may increase your height and hand size.  Height via the twisting forces that are applied by throwing a proper punch causing shearing forces on the spine thereby increasing periosteal width.  Hand size as a result of shearing forces along the carpal bones.

Pitching-It's been documented that a pitchers throwing arm is about three inches longer than his other arm. Pitching puts spiral forces on the bone. Now people have retorted that having one longer arm than the other may cause people to be better pitchers but since I don't have access to pitchers before and after there's nothing I can do to prove it. But say that pitching can improve height, if you can get that significant denaturing of bone with the spiral forces of pitching imagine how much you can get if you add the other three forces.

Chiropractor/Rolfing-I have done both of these and each resulted in a temporary half an inch height gain.  It went away very quickly however and both can be expensive.  Self performed chiropractic and massage techniques tend to be less effective.  They both work as a result of increasing body alignment.  They are only temporary measures and don't significantly affect the bone length.

Sky of easyheight.com's method of causing microfractures via sprinting with ankle weights and then sitting with ankle weights-I think this method is correct in theory but sitting with ankle weights does not provide enough distraction(stretching) forces on the bone.  The theory of stretching the bone coupled with microfractures is what I have based the majority of my height increase research on.  The best way to stretch the bone is to put a weight at the end of the bone to stretch the bone throughout it's length not to sit with ankle weights.

Growth Hormone-An increase in growth hormone levels in children without a growth hormone mutation has only been shown to increase growth rate not final bone length.  Growth Hormone may effectively increase periosteal width however without any growth hormone mutation.

Diet-This is a whole bag of worms.  Certain proteins are necessary for growth which may explain why in certain decades individuals were shorter.  Growth hormone in meat products may result in individuals with longer torsos due to an increase in bone width(bone width increases by the same mechanism as bone length in short and irregular bones).  Chemical factors affect bone growth but chemical factors that could increase height are subject to negative feedback mechanisms whereas mechanical factors that could increase height are better able to get around negative feedback(if a bone is broken it's going to heal negative feedback or no negative feedback).  Chemical factors that reduce height are not as susceptible to positive feedback mechanisms so it may be worthwhile to make sure you have all your nutrition in check.

Tacrolimus(FK506)

Tacrolimus is available by prescription only.

Predominant promotion by tacrolimus of chondrogenic differentiation to proliferating chondrocytes.

"Tacrolimus (FK506) has been used as a therapeutic drug beneficial for the treatment of rheumatoid arthritis in humans. In this study, we investigated the effects of FK506 on cellular differentiation in cultured chondrogenic cells. Culture with FK506 led to a significant and concentration-dependent increase in Alcian blue staining for matrix proteoglycan at 0.1 to 1,000 ng/ml, but not in alkaline phosphatase (ALP) activity, in ATDC5 cells, a mouse pre-chondrogenic cell line, cultured for 7 to 28 days, while the non-steroidal anti-inflammatory drug indomethacin significantly decreased Alcian blue staining in a concentration-dependent manner, without altering ALP activity. FK506 significantly increased the expression of mRNA for both type II and type X collagen, but not for osteopontin, in ATDC5 cells. Similar promotion was seen in chondrogenic differentiation in both mouse metatarsals and chondrocytes cultured with FK506. However, FK506 failed to significantly affect transcriptional activity of the reporter construct for either sry-type HMG box 9 (Sox9) or runt-related transcription factor-2 (Runx2), which are both transcription factors responsible for chondrocytic maturation as a master regulator. These results suggest that FK506 may predominantly promote cellular differentiation into proliferating chondrocytes through a mechanism not relevant to the transactivation by either Sox9 or Runx2 in chondrogenic cells."

"FK506 preferentially inhibits the production of inflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β, through suppression of T-cell activation in human peripheral blood mononuclear cells, without markedly affecting proliferation and differentiation of bone marrow cells"

"The immunosuppressant drugs FK506 and cyclosporine A inhibit the activity of the ubiquitously expressed Ca2+/calmodulin–dependent protein phosphatase, calcineurin, through an interaction between the calcineurin subunit A and different immunophilins such as FK506-binding protein for FK506 and cyclophilins for cyclosporine A, which consequently leads to the inhibition of calcineurin-mediated dephosphorylation of the nuclear factor of activated T cells in T-lymphocyte cells"

"In chondrogenic ATDC5 cells, chondrogenic differentiation is promoted by FK506, but not by cyclosporine A, in terms of matrix proteoglycan production and type II collagen (Col II) expression."

Insulin increased proteoglycan matrix accumulation by ATDC5 cells.

"FK506 would predominantly promote cellular differentiation from pre-chondrocytes to proliferating chondrocytes, without markedly altering that from proliferating to hypertrophic chondrocytes or from hypertrophic to calcified chondrocytes, in cultured metatarsals ex vivo." See Figure 9 for a great graphic explaining the different stages of chondrogenic differentiation.

The immunosuppressant FK506 promotes development of the chondrogenic phenotype in human synovial stromal cells via modulation of the Smad signaling pathway.

"hSSCs were isolated from synovium of the knee joint and 2x10(5) cells were subjected to pellet culture in chondrogenic culture medium for 3 weeks with or without growth factors [bone morphogenetic protein 2 (BMP2) or transforming growth factor beta(1) (TGFbeta(1))] and +/- addition of FK506 in chondrogenic culture media was evaluated. Chondrogenesis was assessed by the size of the pellet, the production of proteoglycans, and messenger RNA (mRNA) levels for chondrogenic markers. Furthermore, levels and intracellular location of phosphorylated Smad proteins related to BMP signaling and TGFbeta signaling were evaluated following exposure to FK506.

FK506 enhanced the differentiation of hSSCs toward a chondrogenic phenotype in a dose-dependent manner associated with increases in glycosaminoglycan synthesis and increased mRNA levels for chondrogenic marker genes. Additionally, FK506 further enhanced chondrogenesis of synovial stromal cells (SSCs) induced by BMP2 and TGFbeta(1), also in a dose-dependent manner. Notably, phosphorylation of Smad1/5/8 and Smad3 was significantly increased by FK506. Also, the ratio of nuclear translocation to cytoplasmic levels of phosphorylated Smad1/5/8 and Smad3 were increased following exposure of SSCs to FK506. Moreover, inhibition of Smad signaling significantly abrogated FK506-induced chondrogenic differentiation of SSCs."

"The immunosuppressive effect of FK506 is believed to be related to its ability to inhibit calcineurin, an enzyme involved in the activation of the nuclear factor of activated T cells (NFAT)"<-inhibiting calcineurin may help make you taller.  Although some NFATs seem to pro-chondrogenic whereas others are anti-.

"SSCs were isolated from human synovium which was harvested from the knee joint of four patients during arthroscopic surgery"<-Ages 19 to 57.

"To assess chondrogenic differentiation, a pellet culture system was used24. Approximately 2 × 105 cells (passages 3–5) were placed in a 15-ml polypropylene tube, and centrifuged at 500g for 10 min. The pellets were cultured at 37°C with 5% CO2 in 500 μl of chondrogenic culture medium that contained HG-DMEM with 10% FBS, supplemented with 50 μg/ml ascorbate-2-phosphate, 100 μg/ml sodium pyruvate, and 50 mg/ml insulin, transferrin, and selenious acid (ITS) + Premix"

Inhibition of the ERK, PI3K, and the p38 pathways did not affect Tacrolimus' enhancement of chondrogenesis.

TRPV4

Human skeletal dysplasia caused by a constitutive activated transient receptor potential vanilloid 4 (TRPV4) cation channel mutation.

"The transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, is expressed in a broad range of tissues where it participates in the generation of Ca2+ signals and/or depolarization of the membrane potential. Regulation of TRPV4 abundance at the cell surface is critical for osmo- and mechanotransduction. Defects in TRPV4 are the cause of several human diseases, including brachyolmia type 3; also known as brachyrachia or spondylometaphyseal dysplasia Kozlowski type, and metatropic dysplasia; also called metatropic dwarfism or parastremmatic dwarfism. These bone dysplasia mutants are characterized by severe dwarfism, kyphoscoliosis, distortion and bowing of the extremities, and contractures of the large joints. These diseases are characterized by a combination of decreased bone density, bowing of the long bones, platyspondyly, and striking irregularities of endochondral ossification with areas of calcific stippling and streaking in radiolucent epiphyses, metaphyses, and apophyses. We emphasize how the constitutive active TRPV4 mutant affects endochondral ossification with a reduced number of hypertrophic chondrocytes and the presence of cartilage islands within the zone of primary mineralization."

I couldn't find this whole paper anywhere.

Why it is necessary to couple muscle building with bone growth in many grow taller methods

For several methods to build bone and the methods applicable for growing taller it's almost impossible to build up bone without building some muscle up as well. It's possible, to provide the forces needed to build bone without weight via "tapping"(hitting the bone with weights or a hammer) or limb lengthening surgery but then you aren't using all the possible forces on bone you can to cause microfractures(compression, strain, spiral, and impact). And in any case many of the bones are "blocked" by muscle in order to tap the bone you also have to tap the muscle as well(tapping is a bit of an underhyperbole as you want to tap as hard as you can without causing nerve damage).

Now, via Lateral Synovial Joint Loading you are putting load on the ends of the bones lateral to the joint area to provide a stretching force along the entire bone.  But, you need muscle in order to control the weight that you're using to provide the downward forces or you need muscle to help protect your tendons and ligaments from injury if it's an external force providing a stretch with downward pressure on the ends of the bones.

In addition, it is also possible to grow taller via an increase in periosteal width in the short and irregular bones.  You need to use weight to create the shearing forces on the short and irregular bones to increase the periosteal width and you need muscle to stabilize your body so you don't, for example, herniate a disc due to a lack of support.

Also, another method for growing taller is via knee microfracture(and microfracture in other areas) in order to release stem cells that exist within bone marrow that stimulate cartilage growth when provided with blood.  The more muscle you have the more force you can generate to create knee microfractures.

In regards to nerve damage, yes their are nerves that are inside the bone but it doesn't necessarily mean you are damaging them. Imagine a wall with electrical wires inside of it, if you bang on the wall you only hurt the wall and not the electrical cables inside of it. The skeletal tissue of the bone will absorb the impact. It's important that you do any tapping yourself, the punch that hurts the most is the one you don't see coming as they say and if you tap your own bones you learn how hard you can tap safely.

Let's address two possible ways to provide forces on bone: Strain fractures via hanging and compression fractures by holding a weight on your back (i.e. in a squat position). If you try to take a weight that you can't handle the golgi tendon will send a signal to your muscle telling it to relax(this is the basis behind massage) and you will not be able to do the "exercise" for very long and you won't cause very many microfractures. You need to do an exercise with a weight that your body will allow you to handle(sometimes this can be quite heavy) so you can continue to progress in weight and cause more microfractures which are necessary to grow via the articular cartilage method and may be necessary to grow taller via distraction forces on the long bones.

We want to try to maximize the forces we place on the bone when we perform exercises designed to increase their size. So, for example when he hang we should also twirl around which generates spiral and shearing forces on the irregular bones of the spine. And, when we perform a walkout in the squat rack to generate compression forces on the irregular bones we should also stomp to provide additional impact and shearing forces on the knee and the bones of the spine.

If your muscles aren't strong enough for the golgi tendon to allow them to go, you'll only be able to perform the exercises for 2 to 3 seconds rather than the perhaps 20 if your muscles(and CNS) are strong enough to allow the exercises to be fully performed. In addition, your muscles(and CNS) won't adapt and you won't be able to move on to higher weights.

Muscular conditioning is also important. You need to train frequently enough so your muscles adapt and are harder to damage thus more of the forces you want can be targeted to the bone and less to the muscle. We want inclusion of bone not exclusion of muscle. There is no such thing as an "isolation" exercise.

We are just trying to specialize our training to gain height rather than excluding forces that damage the muscle entirely.

It's not an LSJL lengthening study but it still provides insights into LSJL and it might help provide more insight into how to perform LSJL.  Although the loads are axial loads rather than lateral loads.  Also, remember that providing pressure on the muscle increases hydrostatic pressure in the bone.

Mechanical loading, damping, and load-driven bone formation in mouse tibiae.
"Loading can activate anabolic genes.  We investigated the damping capacity of bone, joint tissue, muscle, and skin using a mouse hindlimb model of enhanced loading in conjunction with finite element modeling to model bone curvature. Our hypothesis was that loads were primarily absorbed by the joints and muscle tissue, but that bone also contributed to damping through its compression and natural bending. To test this hypothesis, fresh mouse distal lower limb segments were cyclically loaded in axial compression in sequential bouts, with each subsequent bout having less surrounding tissue. A finite element model was generated to model effects of bone curvature in silico. Two damping-related parameters (phase shift angle and energy loss) were determined from the output of the loading experiments. Interestingly, the experimental results revealed that the knee joint contributed to the largest portion of the damping capacity of the limb, and bone itself accounted for approximately 38% of the total phase shift angle. Computational results showed that normal bone curvature enhanced the damping capacity of the bone by approximately 40%, and the damping effect grew at an accelerated pace as curvature was increased. Although structural curvature reduces critical loads for buckling in beam theory, evolution apparently favors maintaining curvature in the tibia. Histomorphometric analysis of the tibia revealed that in response to axial loading, bone formation was significantly enhanced in the regions that were predicted to receive a curvature-induced bending moment[maybe there's less damping effect for lateral rather than axial loads which could be why lateral loads induce different gene expression]."

"Damping is the potential of the whole musculoskeletal system to effectively dissipate energy associated with loading."

"Loads of 5 N at 0.5, 2, and 20 Hz were applied to this FE model for 10 s"

"The ratio between the phase shift angle of stage V (bone alone) and stage I (intact hindlimb) demonstrated the influence of the tibia alone. This ratio was 43% at 0.5 Hz, 38% at 2 Hz, and 28% at 20 Hz. Removal of the ankle, skin, and muscle exhibited the smallest effects on phase shift angle, contributing to approximately 14% of the total phase shift at each frequency."<-So most of the load absorption is by the bone.

"While similar trends in energy dissipation are seen in response to 2 and 20 Hz waveforms, the magnitude of the energy loss in each sample configuration decreased monotonously as the frequency was increased."

"Skin was found to contribute to approximately 12% of the total dissipated energy at all frequencies, and the remaining energy was lost with removal of the muscle and foot."

"it has been proposed that the curved shape of long bones, while reducing overall axial strength, increases predictability and better distributes stresses within the bone during loading"<-maybe axial loading  increases bone curvature thereby reducing height?

The role of muscle cells in regulating cartilage matrix production.

"We tested the hypothesis that muscle cells directly regulate cartilage matrix production by analyzing chondrocytes cocultured with muscle cells in 2D or 3D conditions. We found that chondrocytes cultured with C2C12 muscle cells exhibited enhanced alcian blue staining and elevated expression of collagen II and collagen IX proteins. Although nonmuscle cells did not promote cartilage matrix production, converting them into muscle cells enhanced their pro-chondrogenic activity. Furthermore, muscle cell-conditioned medium led to increased cartilage matrix production, suggesting that muscle cells secrete pro-chondrogenic factors. Taken together, our study suggests that muscle cells may play an important role in regulating cartilage gene expression."

"Mouse mutants that lack muscle-specific proteins such as dystrophin/utrophin or myogenin also exhibited skeletal abnormalities such as a curved spine or a reduced size of the skeleton"

"C2C12 muscle cell have an elongated, fibroblast-like morphology, while RCS chondrocytes are rounded in shape. When co-cultured, DiI-positive cells still have the fibroblast-like morphology, while all unlabeled cells maintain a round morphology. This indicates that muscle cells and chondrocytes do not change their morphology upon co-culturing"

"upon co-culturing, muscle cells herd the chondrocytes into clusters and the chondrocytes assume a rounder phenotype. Furthermore, chondrocytes (RCS) co-cultured with mouse C2C12 muscle cells expressed higher levels of collagen II and collagen IX proteins"

"muscle secretes a variety of growth factors or cytokines that can be carried away by blood or interstitial fluid. Among them is the known pro-chondrogenic factor IGF-I"

Bone, muscle, and physical activity: structural equation modeling of relationships and genetic influence with age.

"Quantitative trait locus (QTL) analysis was used to identify regions of chromosomes that simultaneously influenced skeletal mechanics, muscle mass, and/or activity-related behaviors in young and aged B6xD2 second-generation (F(2)) mice of both sexes. SEM was used to further study relationships among select QTLs, bone mechanics, muscle mass, and measures of activity. The SEM approach provided the means to numerically decouple the musculoskeletal effects of mechanical loading from the effects of other physiological processes involved in locomotion and physical activity. Muscle mass was a better predictor of bone mechanics in young females, whereas mechanical loading was a better predictor of bone mechanics in older females. An activity-induced loading factor positively predicted the mechanical behavior of hindlimb bones in older males; contrarily, load-free locomotion (i.e., the remaining effects after removing the effects of loading) negatively predicted bone performance. QTLs on chromosomes 4, 7, and 9 seem to exert some of their influence on bone through actions consistent with Wolff's Law."

Here's a diagram of how muscle may interact with bone:

Although body length is used as an independent predictor.

MyoD1 and IGF1R are two genes mentioned as being associated with muscle and bone.

Snail1/Snail2

METHODS OF IDENTIFYING AND USING SNAIL1 INHIBITORY COMPOUNDS IN CHONDRODYSPLASIA TREATMENT AND PREPARATION OF PHARMACEUTICAL COMPOSITIONS

"Snail1[or Snai1] gene contributes to FGFR3 receptor signal transduction, which contributes to chondrodysplasias (achondroplasia (ACH), thanatophoric dysplasia (TD) and hypochondroplasia (HCH)). The exemplary embodiments identify Snail1 as a therapeutic and diagnostic target for chondrodysplasia, as well as the use of inhibitors thereof as drugs for the treatment of these diseases."

"Chondrocytes and osteoblasts have a mesenchymal origin, whereas osteoclasts arise from the haematopoietic lineage."

"the long-bone shortening phenotype [of being FGFR3 transgenic] is due to a disorganization and shortening of the proliferative chondrocyte columns and to delayed differentiation. Defects in the proliferative chondrocytes are due to the activation of Stat1, which is responsible for the induction of cell cycle inhibitor p21, and delayed differentiation is due to the activation of the MAPK signaling cascade, which causes a reduction in the area of hypertrophic chondrocytes, in both animal and human models. On the contrary, the de-activation of Fgfr3 in mice causes prolonged endochondral growth, resulting in a "long-bone" phenotype, which is accompanied by an expansion of the proliferative chondrocyte region in the growth plate. All these data give FGFR3 a significant role as a negative regulator of the proliferation of chondrocytes. This inhibition of proliferation by the FGF pathway is unique to chondrocytes and is mediated by transcription factor STAT1, which increases the expression of cell cycle inhibitor p21, the final agent responsible for the interruption of the proliferation induced by this signaling pathway. [p21] levels can be considered to be a reflection of the activation of the FGFR3-mediated signaling pathway in the growth plate. "

"Snail1 [is] a direct repressor of type II collagen, which is characteristic of proliferative chondrocytes, and disappears when the latter cease to proliferate and differentiate into hypertrophic chondrocytes, the cell population that expresses type X collagen. In a completely independent context, it was observed that the presence of Snail attenuated the proliferation of epithelial cells in culture and evolved with an increase in p21 levels and an increase in the phosphorylation of ERK1 and ERK2."

"The use of natiuretic peptide CNP inhibits the FGF-mediated MAPK activation in the growth plate"

"Snail1 is sufficient for FGFR3 signaling in the bone."

"Exemplary inhibitory compounds include, without limitation, antisense oligonucleotides described in, without limitation, US Patent Publication No. US20060003956 and Kajita et al., 2004 or siRNAs that inhibit the expression of Snail1 such as those described in without limitation, Peinado et al., 2005 and Tripathi et al., 2005 all of which are incorporated by reference herein. Additionally, any published nucleotide sequences or those published in the future that inhibit the expression of Snail1 are incorporated as embodiments herein, as potentially useful therapeutic compounds for the preparation of drugs designed to treat a chondrodysplasia process. Gene inhibition techniques and, more specifically, transport of compounds including, without limitation, antisense oligonucleotides, siRNA, ribozymes or aptamers--can be performed using, without limitation, liposomes, nanoparticles or other carriers that increase the success rate of the transfer to the interior of the cell, in one exemplary embodiment the cell nucleus (see, without limitation, Lu and Woodle, 2005 and Hawker and Wooley, 2005 which are incorporated by reference herein). In principle, Snail1 mRNA translation inhibitors can be used which bind both to the encoding region and/or the non-encoding region, for example, in front of the 3' non-encoding area. "


Compensatory regulation of the Snai1 and Snai2 genes during chondrogenesis.

"Several genes involved in cartilage and bone development have been identified as target genes for the Snail family of zinc finger transcriptional repressors, and a gain-of-function study has demonstrated that upregulation of Snai1 activity in mouse long bones caused a reduction in bone length. The Snai1 and Snai2[also known as slug] genes function redundantly during embryonic long bone development in mice. Deletion of the Snai2 gene, or limb bud-specific conditional deletion of the Snai1 gene, did not result in obvious defects in the skeleton. However, limb bud-specific Snai1 deletion on a Snai2 null genetic background resulted in substantial defects in the long bones of the limbs. Long bones of the Snai1/Snai2 double mutants exhibited defects in chondrocyte morphology and organization, inhibited trabecular bone formation and delayed ossification. Chondrocyte proliferation was markedly reduced, and transcript levels of genes encoding cell cycle regulators, such as p21(Waf1/Cip1) , were strikingly upregulated in the Snai1/Snai2 double mutants, suggesting that during chondrogenesis Snail family proteins act to control cell proliferation by mediating expression of cell cycle regulators. Snai2 transcript levels were increased in Snai1 mutant femurs, while Snai1 transcript levels were increased in Snai2 mutant femurs. In addition, in the mutant femurs the Snai1 and Snai2 genes compensated for each other's loss not only quantitatively, but also by expanding their expression into the other genes' normal expression domains. The Snai1 and Snai2 genes transcriptionally compensate temporally, spatially, and quantitatively for each other's loss, and demonstrate an essential role for Snail family genes during chondrogenesis in mice."

"mean bone length of femurs of the Snai1/Snai2 DM embryos was reduced approximately 20%, whether the measurements were taken of total bone length, or the length of trabecular bone"

"in Snai1/Snai2 DM growth plates the pattern of well-aligned columnar chondrocytes was disorganized, and chondrocyte morphology was altered. Proliferating chondrocytes in Snai1/Snai2 DM growth plates were more compact, and exhibited an elliptical shape with a higher chondrocyte to lacuna ratio"

"statistically significant increase in the length of the hypertrophic chondrocyte zone in Snai1/Snai2 DM growth plates"

"Snai1 and Snai2 genes are required to maintain the high rate of chondrocyte proliferation in the rapidly growing long bone"

"Cdkn1a RNA expression was approximately six-fold higher in Snai1/Snai2 DM femurs than in littermate controls"

"several markers were upregulated in Snai1/Snai2 DM femurs, including Col1a1, Col2a1, Col10a1, Sox9, and Acan"<-all these genes were upregulated by LSJL.

"genes encoding the matrix metalloproteases Mmp9 and Mmp13 were downregulated"

"Progression into chondrocyte prehypertropy and hypertrophy appeared to be delayed in Snai1/Snai2 DM growth plates. At E14.5, expression of both Col10a1, a hypertrophic chondrocyte marker, and Indian hedgehog (Ihh), a prehypertrophic chondrocyte marker, was markedly delayed in double mutant femurs"

"Blood vessel formation and penetration into the trabecular region of the femur was observed in both double mutant and controls"

"Despite the increase in Trp53 expression [in double mutants], we did not observe increased chondrocyte cell death in these embryos"

Snai1 mutant knockout mice had a slight non-signficiant increase in bone length.  At E16.5, Snai mutant knockout mice had a slight non-significant increase in Col10A1 and MMP9 levels.

For Height don't take Ammonium Chloride?

The Effect of Excessive Acid Feeding on Bone

"The purpose of the present study was to examine the effect of excessive ammonium chloride administration and/or low calcium diet on bone in adult rats."

"Male Sprague-Dawley rats were allowed to grow on regular laboratory diet and tap water to an average weight of 550 g."<-Whether these rats were post growth plate senescence is unclear.

"1. Regular laboratory diet and distilled water.
2. Regular laboratory diet and 2 % NH~Cl.
3. Special low calcium diet and distilled water.
4. Special low calcium diet and 2 % NH~Cl."

"The animals were allowed to consume the diet and the drinking fluid ad lib and were killed after 6 months."

"When compared with the baseline control group, the bones of the experimental animals in all four groups are seen to have grown approximately 0.2 cm in length."<-Group 1 gained the most height.  Group 4 actually gained the second most height so maybe low calcium diet and ammonium chloride have a synergestic effect.

Grow in Height with Amylin and adrenomedullin?

Systemic administration of amylin increases bone mass, linear growth, and adiposity in adult male mice.

"Amylin is a peptide hormone cosecreted with insulin from the pancreatic beta-cells that can act as an osteoblast mitogen. Adult male mice[age 40-50 days weighing 25 to 32 grams] were given daily subcutaneous injections of amylin (10.5 microgram) or vehicle (n = 20 in each group) for 4 wk. Histomorphometric indices of bone formation increased 30-100% in the amylin-treated group, whereas resorption indices were reduced by approximately 70% (P < 0.005 for all indices). Total bone volume in the proximal tibia was 13.5 +/- 1.4% in control animals and 23.0 +/- 2.0% in those receiving amylin (P = 0.0005). Cortical width, tibial growth plate width[by about double], tibial length[by about 5-10%], body weight, and fat mass were all increased in the amylin-treated group. It is concluded that systemic administration of amylin increases skeletal mass and linear bone growth."

Amylin alters renal calcium handling and increases circulating IGF-1 levels.

"[Chondrocytes] show increased thymidine incorporation and increased cell numbers after treatment for 24 h with amylin in concentrations of 1 × 10−10 M and greater"

Amylin and adrenomedullin: novel regulators of bone growth.

"The peptide hormones amylin and adrenomedullin [are] known to circulate at picomolar concentrations. Adrenomedullin is also produced locally in bone. Amylin and adrenomedullin are related peptides with some homology to both calcitonin and calcitonin gene-related peptide."

"Both amylin and adrenomedullin have also been found to act on chondrocytes, stimulating their proliferation in culture and increasing tibial growth plate thickness when administered systemically to adult mice."

Relationships between fat and bone.

"Fat mass [is associated with] with the secretion of bone active hormones from the pancreatic beta cell (including insulin, amylin, and preptin), and the secretion of bone active hormones (e.g., estrogens and leptin) from the adipocyte."

"Amylin is co-secreted with insulin and potently inhibits bone resorption"

"deletion of GLP-2 receptor leads to marked skeletal deficits in growing mice"

Adrenomedullin(22-52) combats inflammation and prevents systemic bone loss in murine collagen-induced arthritis.

"DBA/1 mice with collagen-induced arthritis (CIA) were treated with 1.2 μg/gm adrenomedullin(22-52) , adrenomedullin, or saline at arthritis onset."

"Tumor necrosis factor α, interleukin-6 (IL-6), and IL-17A levels were significantly decreased in the joints of mice with CIA treated with adrenomedullin or adrenomedullin(22-52) as compared to controls, whereas IL-4 and IL-10 levels were increased. Adrenomedullin(22-52) was more effective than adrenomedullin in modulating cytokine content and enhanced Treg cell function without changing Treg cell expression compared to controls. Adrenomedullin receptor binding and transcriptional adrenomedullin receptor expression were markedly increased in joints from controls, whereas adrenomedullin receptor binding was considerably decreased in treated animals. Mice with CIA treated with adrenomedullin or adrenomedullin(22-52) had considerably fewer apoptotic chondrocytes and diminished cartilage degradation. Adrenomedullin(22-52) completely prevented systemic bone loss by preserving osteoblastic activity, but without changes in osteoclastic activity."

" Murine and human chondrocytes express adrenomedullin receptors"

"Adrenomedullin receptors are composed of the calcitonin-like receptor (CLR). CLR is a 7-transmembrane G protein associated with 1 of 3 single membrane–spanning receptor activity–modifying proteins (RAMPs), namely, RAMP-1, RAMP-2, or RAMP-3."

Expression of adrenomedullin and its receptor by chondrocyte phenotype cells.

"[We] investigate the secretion of adrenomedullin (AM) by chondrocyte phenotype cells and whether or not AM effects this proliferation in a cAMP-dependent fashion. Chondrocyte phenotype cells expressed AM and the AM receptor, and secreted high concentration of AM into the culture medium. When added to cultures, AM increased the intracellular cAMP level and decreased the number of these cells in a similar concentration-dependent fashion. Addition of forskolin and dibutyryl-cAMP caused a significant decrease in the number of these cells. Furthermore, the effect of AM was inhibited by a cAMP-dependent protein kinase A inhibitor (H89). The present findings indicate that AM has an autocrine/paracrine type of anti-proliferative effect on these cells mediated via a cAMP-dependent pathway and raise the possibility that AM plays a role in the local modulation of a process of de-differentiation by culturing"

So Adrenomedullin likely helps further the differentiation of chondrocyte cells.

"When chondrocytes are cultured, there is prompt down-regulation of expression of cartilage-specific genes, including those encoding type-2, -9, and -11 collagens and aggrecan{all up in LSJL}, as well as concomitant initiation or up-regulation of expression of fibroblast-associated genes, including those for type-1{up}, -3{up}, and -5{up} collagens and versican{up}"

"Cyclic AMP acts as a second messenger in various tissues, including fibroblasts and chondrocytes, and in cultured fibroblasts and chondrocytes, cAMP analogs decrease thymidine incorporation into DNA"

"the addition of forskolin and dibutyryl-cAMP caused a significant decrease in cell population. AM activity is likely to be primarily mediated via the cAMP pathway and involves inhibition of the mitogen-activated protein kinase cascade"

"H89, a PKA inhibitor, blocked the effects of AM on chondrocyte phenotype cells"

Effects of amylin and adrenomedullin on the skeleton.

"Amylin is secreted following eating and may direct calcium and protein absorbed from the meal into new bone synthesis. Amylin circulates in high concentrations in obese individuals, and might contribute to the association between bone mass and fat mass. "

"[Amylin is] co-secreted with insulin from the pancreatic B-cell"

"adrenomedullin [is] in maturing cartilage, hypertrophic cartilage and in osteoblasts of the developing bone."

Amylin increases tibial length and growth plate width.

The logical fallacies about growing taller

For approximately six years, I've studied how to grow taller. I've encountered several logical fallacies from people who wanted to discourage me from my efforts..

1) If it was possible to grow taller someone would've discovered it by now.

This argument only works for goals that have a low opportunity cost(i.e. you can gain abs of steel by using this machine for thirty minutes once a month!). If gaining height by non-surgical means takes a lot of effort, then it is entirely possible that basketball coaches, volleyball coaches, et. al. have not discovered it yet. After all, why have your players do something fatiguing when you could get already tall people and teach them the art of basketball?

The research on bone is inadequate. Research takes years to perform and they don't usually test for something they're not looking for. I used to work part time at a lab and the studies were highly complex to determine something so simple. A researchers hypothesis has to be simple to eliminate confounding variables. You have to get grant approval for funding, etc. The scientific method is the ultimate piece of red tape and the equipment used to study bones is extremely expensive. Also, most studies on bone are tested on mice and last I checked mice can't do the things that humans can do like hanging or deadlifts. At the rate scientists work at, it could take years for them to discover a non-surgical way of growing taller.

2) The growth plates fuse after puberty therefore it is not possible to grow taller.

Just because growing taller is no longer possible one way does not necessarily eliminate the ability to grow taller another way. We already know this to be the case via the surgical method. The key to growing taller involves microfractures and distraction(stretching) forces. The way to cause these microfractures is via spiral, impact, strain, and compression forces.

Microfractures can propagate as a result of distraction forces on the bone. These microfractures help the bone maintain it's distracted state and over time result in a long bone.

In addition, short and irregular bones are completely covered with periosteum and it is possible to increase periosteal width via exercise and even growth hormone.


3) If it were possible to gain height via specialized weight training why aren't bodybuilders super tall?

First, a Bone Mineral Density scan measures a bones size, mineral content, and porosity so there is no way to tell in vivo(for an alive subject) that a bone has grown bigger. How do you know whether it's not an increase in mineral content or a decrease in porosity making the BMD bigger(both of which do not affect height)?

Secondly, at the doctors office they do not do a thorough test in height. The procedure is designed to be quick and easy. That's the basis of the BMI after all so they don't have to do accurate body fat tests. Any small increase in height would be tossed aside as measurement error. Height is not the pentultimate indicator of skeletal size. It is possible for the skeleton to grow bigger without a large increase in height. We are three dimensional objects after all.

Third, the way people weight train may not be the optimal way for gaining height. The way most people weight train may not provide heavy enough spiral, compression, strain, or impact forces to get a large increase in bone size. The way most people weight train produces a larger load on the muscle but not a very large load on the bone. Of course, this should still produce a bigger bone(the same way that say a runner gains more muscle but not nearly as much as a bodybuilder) but not at a rate large enough to be unexplainable by measurement error.

Fourth, scientists are biased against testing their theories on weightlifters. Scientists are not willing to risk the perceived injury risk that heavy weightlifting may cause.

CXCR4

CXCR4 is involved in cell homing.

Stimulation of chondrocyte hypertrophy by chemokine stromal cell-derived factor 1 in the chondro-osseous junction during endochondral bone formation.

"G-protein coupled receptor CXCR4 is predominantly expressed in hypertrophic chondrocytes, while its ligand, chemokine stromal cell-derived factor 1 (SDF-1) is expressed in the bone marrow adjacent to hypertrophic chondrocytes. Thus, they are expressed in a complementary pattern in the chondro-osseous junction of the growth plate. Transfection of a CXCR4 cDNA into pre-hypertrophic chondrocytes results in a dose-dependent increase of hypertrophic markers including Runx2, Col X, and MMP-13 in response to SDF-1 treatment. In organ culture SDF-1 infiltrates cartilage and accelerates growth plate hypertrophy. Furthermore, a continuous infusion of SDF-1 into the rabbit proximal tibial physis results in early physeal closure, which is accompanied by a transient elevation of type X collagen expression. Blocking SDF-1/CXCR4 interaction suppresses the expression of Runx2. Thus, interaction of SDF-1 and CXCR4 is required for Runx2 expression. Interestingly, knocking down Runx2 gene expression results in a decrease of CXCR4 mRNA levels in hypertrophic chondrocytes. This suggests a positive feedback loop of stimulation of chondrocyte hypertrophy by SDF-1/CXCR4, which is mediated by Runx2."

"degradation of hypertrophic cartilage requires the presence of bone marrow"

Epiphyseal Plate Vascularization

Epiphyseal plate vascularization refers to the post hypertrophic stage of the growth plate where the growth plate is invaded by blood vessels before the ossification zone forms.


Matrix vesicles: structure, composition, formation and function in calcification.

"Matrix vesicles (MVs) induce calcification during endochondral bone formation. MV proteins, enzymes, receptors, transporters, regulators, lipids and electrolytes are detailed. MV formation is considered from both structural and biochemical perspectives. Confocal imaging of Ca(2+) and H(+) were used to depict how living chondrocytes form MVs. coordinated mitochondrial Ca(2+) and Pi metabolism produce MVs containing a nucleational complex (NC) of amorphous calcium phosphate, phosphatidylserine and annexin A5--all critical to the mechanism of mineral nucleation. Reconstitution of the NC and modeling with unilamellar vesicles reveal how the NC transforms into octacalcium phosphate, regulated by Mg(2+), Zn(2+) and annexin A5. Extravasation of intravesicular mineral is mediated by phospholipases and tissue-nonspecific alkaline phosphatase (TNAP). In the extravesicular matrix, hydroxyapatite crystal propagation is enhanced by cartilage collagens and TNAP, which destroys inhibitory PPi, and by metalloproteases that degrade proteoglycans."

"Endochondral calcification is a primary, but provisional process; its end product, calcified cartilage, is more heavily calcified, but is mechanically weaker than true bone. It is destined to be largely replaced by cancellous bone. Endochondral calcification is mediated by growth plate chondrocytes; it is a rapid de novo process that begins with Ca2+ and Pi ions in solution. It involves overcoming of a sizeable nucleation barrier that requires the direct mediation of cellular metabolic activity. It utilizes extracellular vesicles and ends with formation of large amounts of microcrystalline, Ca2+-deficient, acid-phosphate-rich apatite deposits embedded in a proteoglycan and cartilage-specific collage-rich matrix."

"Type VI collagen receptors on the chondrocyte surface (NG2-proteoglycan) act as transducers to mitochondria within the cells, affecting their permeation transition pore for Ca2+" LSJL upregulates Col6a1,Col6a2, and Col6a3.  And downregulates another form of Col6a2.  Col6 is involved in matrix vesicle formation.

"Type X collagen had a small inhibitory effect on the onset and rate of mineral formation; however it significantly extended the length of the "rapid formation" period, thereby altering the kinetics of the overall amount of mineral formation"

Anxa5, 6, 1, 11, 4, and 2 are associated with MVs.

"AnxA5 greatly accelerates the nucleational activity of the acidic phospholipid-Ca2+-Pi complexes present in the nucleational core that triggers de novo calcium phosphate mineral formation in MVs. AnxA5 specifically overcomes the inhibitory effects of Mg2+ that otherwise greatly slow the induction of mineral formation"

"Expression of AnxA6 in cells reduces cell proliferation"

"AnxA2 has been implicated in the detachment of membrane plaques from actin filaments, leading to the formation of cell-surface blebs"

"NPP1 accounts for nearly all the ecto-pyrophosphatase activity of chondrocytes and the release of PPi is thought to provide a means for regulating apatitic mineral formation"

"phospholipid scramblase activity in growth plate chondrocytes [is associated] with TNAP-containing lipid rafts during MV formation"

"In the maturing growth plate chondrocytes, the presence of TRPV4 very likely has an important function in the major osmotic swelling that occurs upon hypertrophy when cell volume expands over 9-fold going from the proliferating to the hypertrophic state. Also of interest are CLIC4 - the Channel 4 intracellular Cl- transporter; this channel protein transports Cl- in exchange for HCO3-, and appears to be important in buffering the internal pH of MVs. The third is VDAC-2, - the Channel 2 voltage-dependent anion-selective channel present in the mitochondrial outer membrane (312-313). This channel may play a part in the mitochondrial permeability transition pore that becomes activated prior to formation of MVs by hypertrophic chondrocytes"

SLC's involved in MV's:

"SLC 16A1{down} [is] a transporter known to transport lactate, pyruvate"

"neutral amino acids (SLC38A3), alanine/serine/cysteine/threonine (SLC1A4){up}, organic cations, e.g. carnitine (SLC22A16), glycine (SLC6A9), Na+-dependent citrate (SLC13A5), Na+-independent cationic amino acids (SLC7A3){down}, and equilibration of nucleosides (SLC29A1). "

"two Zn2+ transporters - SLC39A14 and SLC39A8."

"Pi transporters use an inwardly directed electrochemical Na+ current to support Pi influx. This Na+ gradient, produced by the action of a Na+/K+ ATPase in the plasma membrane, carries Pi into living cells. However in MVs, Na+ levels are as high as that of the extravesicular fluid; thus there is no Na+ gradient to drive Pi uptake."

"Substitution of Li+ or K+ for Na+ had minimal effect on Pi uptake; N-methyl D-glucamine (NMG+) was totally inhibitory, whereas choline+ was clearly stimulatory"

"syntenin binds to syndecan, which in turn binds to type VI collagen in the extracellular matrix, leaving MVs attached at the points of contact"

" phalloidin, an agent that stabilizes actin microfilaments, significantly decreased MV formation. Another drug, colchicine, which destabilizes microtubules, also stimulated the release of MVs."

"both microtubules and microfilaments were involved in the formation of cell processes from which MV were shown to derive"

"accumulated mitochondrial Ca2+ becomes subsequently transferred to MV during growth plate development."

"Accumulation of excessive Ca2+ is known to inhibit mitochondrial respiration "

"Growth plate cartilage has been shown to possess unusually high levels of both cytosolic (20-25 mM) and extracellular (2.2-2.5 mM) Pi"

"cultured tibial growth plate chondrocytes to even modestly elevated levels (3 mM) of extracellular Pi induces 'apoptotic' cell death within 48 h."

"Mitochondrial uptake of Pi is largely accomplished by PiC, an H+/Pi symporter (or Pi/OH- antiporter). PiC is an integral membrane protein, a homodimer that resides in the inner mitochondrial membrane that translocates or exchanges Pi from the cytosol into the mitochondrial matrix"

"PiC is Na+-independent and is responsible for the rapid import of Pi used for ATP synthesis."

"During MV formation, PS-CPLXs can readily form at pH 6.7, conditions like that present in the perimeter of growth plate chondrocytes. In contrast, the ACP component of the nucleational core cannot form at this pH and can occur only after the vesicles are released into the extracellular fluid where the pH (7.6) favors ACP formation.  A key requirement in the formation of PS-CPLX is that excess Pi must be present[which occurs during MV formation] with the lipid before introduction of Ca2+"

"[factors] influencing MV nucleation include: a) the inhibitory effects of Mg2+, Zn2+, and PPi, b) the aforementioned synergistic effects of the MV annexins and PS, and c) the inhibitory effects of PE and other lipid components of the MV membrane."

"MVs are formed by growth plate cells whose cytosolic levels of Mg2+ are much higher than those of Ca2+ . Thus, all PS-Ca2+-Pi complexes formed in vivo incorporate both Mg2+ and Zn2+; in fact analyses of the complexes present in avian MVs have shown that Mg2+ is almost as abundant as Ca2+. Our recent studies have revealed that this level of Mg2+ is profoundly inhibitory to the nucleational activity of PS-Ca2+-Pi complexes, greatly reducing their rate . This inhibitory effect is physiologically important because as noted above rapidly growing species like broiler-strain chickens have such fast longitudinal growth that speedy nucleation is essential for mineral formation to keep pace."

"levels of extravesicular Zn2+ as low as 5 microM are sufficient to totally block MV mineral formation"

" MVs bind tightly to type II and X collagens present in growth plate cartilage"

"during growth plate development the chondrocytes initially require Pi for synthesis of DNA, RNA and nucleotides essential for proliferation. Upon termination of cell division, the nucleotides begin to be degraded; 31P-NMR and chemical analyses reveal that Pi builds up in the cytoplasm. Simultaneously, in situ confocal imaging reveals that the chondrocytes acquire and process Ca2+ at an accelerated pace until in the early hypertrophic zone this climaxes with the cells exfoliating from their plasma membrane the accumulated Ca2+ and Pi as ion-loaded vesicles. This mineral phase, present as a nucleational core within the vesicle lumen, is primarily noncrystalline calcium phosphates complexed with both PS and the annexins, which facilitate the induction of crystalline mineral formation. When MVs are formed by the cells they contain not only Ca2+ and Pi, but also other electrolytes that significantly affect the kinetics of mineral formation; these include Mg2+, Zn2+, and PPi, as well as HCO3-. Production of mineralization-competent MVs enables mineral formation to keep pace with cellular proliferation, preventing development of rickets or osteomalacia. Thus, endochondral bone formatioin involves the provisional calcification of the cartilaginous matrix by chondrocytes - i.e. the formation of calcified cartilage - a more heavily calcified, but mechanically weaker temporary scaffolding that maintains mechanical stability until the truly robust type I collagen-hydroxyapatite composite, bone, is generated by osteoblasts."

MMP2

MMP2 is upregulated by LSJL.  MMP2 is stimulated by GPC3 and GP3 knockout causes overgrowth.


Testican-1, an inhibitor of pro-MMP-2 activation, is expressed in cartilage.

"Testican-1 has been described to be an inhibitor of MT1-MMP and MT3-MMP mediated pro-MMP-2 activation. We studied the expression and localization of testican-1 in human articular cartilage.

Cartilage samples from the medial and lateral tibia plateau were obtained from osteoarthritic patients who underwent joint replacements, and were graded histomorphologically by Mankin score. Testican-1 expression was assessed in RNA isolated directly from cartilage as well as in freshly isolated chondrocytes by reverse transcriptase-polymerase chain reaction (RT-PCR) and quantified by real-time RT-PCR. Testican-1 protein was localized by immunohistochemistry in human osteoarthritic cartilage samples, in human fetal knee joint, and in knees from mice.

Testican-1 mRNA could be detected in cartilage and in freshly isolated chondrocytes both from moderately and from severely damaged osteoarthritic cartilage. In the same donor, expression in chondrocytes from more severely affected regions was decreased compared with chondrocytes from less affected regions. By immunolocalization, testican-1 protein could be detected in chondrocytes predominantly of the superficial and transitional zones. Matrix staining in these zones was greatly reduced in samples from more severely affected osteoarthritic cartilage. A similar distribution was found in the articular cartilage of knees from 7-week-old mice. In addition to articular cartilage, testican-1 was also present in growth plate cartilage.

Testican-1 is a component of cartilage, both of the joint and of the growth plate. Given its activity as an inhibitor of MT1-MMP mediated pro-MMP-2 activation, it is reasonable to speculate that it participates in the regulation of matrix turnover in cartilage."

"Testican-1 is a highly conserved chimeric proteoglycan carrying both chondroitin sulfate and heparan sulfate chains, initially characterized as the precursor of a seminal plasma glycosaminoglycan-bearing peptide"

"testican-1 has been shown to inhibit the cysteine protease cathepsin L, and the heparan sulfate chains present on testican may contribute to the regulation of the collagenolytic activity of cathepsin K"

Testican1 was expressed most strongly in the proliferating zone.

"MMP-2 is an activator of pro-MMP-13"<-MMP13 may actually be beneficial for height growth when expressed in the hypertrophic zone.

Egg Yolk

Effects of Egg Yolk Proteins on the Longitudinal Bone Growth of Adolescent Male Rats

"The bone growth rate was significantly increased by yolk water-soluble protein (YSP, 100 mg/kg) administration for 5 d. The bone morphogenetic protein-2 immunostaining of growth plate was also increased. Considering the results, YSP can be used as a growth-promoting factor."

Egg yolk contains protein, lipid, ash, and carbohydrate.  It has a cholesterol content of 0.1%.  Mice were 3 weeks old.  Yolk water soluble protein promoted growth the most.

Jaoga-Yukmiwon

Effects of Jaoga-Yukmiwon®, a Korean herbal medicine, on chondrocyte proliferation and longitudinal bone growth in adolescent male rats

"The effects of the Korean herbal medicine, Jaoga-Yukmiwon®, on the growth of adolescent rats were investigated in the present study. The proliferation ratio of chondrocytes was calculated from 5-bromo-2′-deoxyuridine incorporation during DNA synthesis. The 5-bromo-2′-deoxyuridine incorporation ratio of the control group was 21.1 ± 3.5%, and Jaoga-Yukmiwon® administration markedly increased the ratio to 34.5 ± 4.6% (p < 0.05). The bone formation rate of longitudinal bone was estimated by labelling with tetracycline, which binds to newly formed bone. The bone formation rate in the tibia of Jaoga-Yukmiwon®-administered rats was increased to 405.1 ± 4.3 µm (p < 0.05) from 292.2 ± 11.8 µm (control value). The height of the growth plate in the administrated rats was increased to 614.6 ± 10.0 µm (p < 0.05) from 552.3 ± 17.1 µm. The bone morphogenetic protein-2 immunostaining in the growth plate was also increased. These results suggest that Jaoga-Yukmiwon® may promote longitudinal bone growth during the developmental period."

"[JY] consists of the alcohol extract of seven medicinal plants, namely Acanthopanacis Cortex (Ja-Oh-Ga in Korean), Rehmanniae Radix Preparat (Suk-Ji-Hwang), Poria (Bok-Ryong), Dioscoreae Rhizoma (San-Yak), Corni Fructus (San-Su-Yu), Cervi Cornu (Nok-Kac) and Ginseng Radix (Go-Ryo-In-Sam). The main component of JY is Acanthopanacis Cortex which has been used for growth disorder in Korean medicine"

"Adolescent male Sprague-Dawley rats (3 weeks of age) were used in the experiment"

"Rats were divided into two groups (n = 8 in each group). The control group was given a daily oral administration (p.o.) of saline and the JY group was given a daily administration of JY (100 mg/kg, p.o.) for 5 consecutive days in each case.  On days 4 and 5 of experiment, all animals were injected
intraperitoneally with 10 mg/kg tetracycline for the measurement of bone growth. On day 6, BrdU
(30 mg/kg) was injected intraperitoneally 1 h before killing to label proliferating cells. One hour after the injection, animals were totally anaesthetized using ether and killed."