Can you grow taller with Lithium Salts?

Previously, I have written about the possibility of Lithium being used as a supplement to help you grow taller during development.  Not much was written about it other than that Lithium inhibits GSK3 beta which inhibits cellular proliferation.  I've been taking Lithium and my Lithium levels are about .26 while effective lithium blood levels are reported to range from 0.5-1.0 mM.  I'm not sure if that benefit is only for the neuronal advantages of Lithium.  Since Lithium increases stem cell proliferation it could also be used to augment LSJL related growth.  Lithium also inhibits Thyroid hormone and I wanted to explore that as well to see if that had any negative impacts on growth.

A role for a lithium-inhibited Golgi nucleotidase in skeletal development and sulfation.

"Sulfation is an important biological process that modulates the function of numerous molecules. It is directly mediated by cytosolic and Golgi sulfotransferases, which use 3'-phosphoadenosine 5'-phosphosulfate to produce sulfated acceptors and 3'-phosphoadenosine 5'-phosphate (PAP). Golgi-resident PAP 3'-phosphatase (gPAPP) is potently inhibited by lithium in vitro. The inactivation of gPAPP in mice led to neonatal lethality, lung abnormalities resembling atelectasis, and dwarfism characterized by aberrant cartilage morphology. The phenotypic similarities of gPAPP mutant mice to chondrodysplastic models harboring mutations within components of the sulfation pathway lead to the discovery of undersulfated chondroitin in the absence of functional enzyme. Loss of gPAPP leads to perturbations in the levels of heparan sulfate species in lung tissue and whole embryos. Clearance of the nucleotide product of sulfotransferases within the Golgi plays an important role in glycosaminoglycan sulfation, provide a unique genetic basis for chondrodysplasia, and define a function for gPAPP in the formation of skeletal elements derived through endochondral ossification."

Remember in the article about FGF, proteoglycan desulfation was a critical component in maximizing height growth.  Sulfated GAGS like chondroitin sulfate consumed like lysosomes which remove waste molecules that can reduce your height growth.  This article says that sulfated chondroitin(and other GAGs) is needed for height.  What's likely is that the ability to remove sulfate chains from GAGs(like Chondroitin) and add new sulfate chains is critical for certain stages of endochondral ossification.  Too much lithium is bad just like other beneficial compounds like BMP-2 or Estrogen etc.  That doesn't mean a little lithium is bad just like a little BMP-2 is bad(it's critical).

"Establishment of the cartilaginous template is a tightly ordered process that involves the function and interplay of: (i) specific transcription factors; (ii) chondrocyte-modulatory ligand pathways; (iii) extracellular matrix (ECM) structural proteins; (iv) core proteins of proteoglycans and their covalently linked glycosaminoglycans (GAGs), such as chondroitin sulfate (CS) chains; and correspondingly (v) gene products essential to the appropriate synthesis and sulfation of GAGs"

"Members of this phosphatase family are functionally inhibited in vitro and in vivo by clinically appropriate doses of lithium "<-Although lower doses of Lithium may not have this effect.

"The extent of gPAPP influence on GAG sulfation likely depends on multiple factors, including: (i) the metabolic load of specific sulfation reactions and thus how the consumption of PAPS by one gSULT may influence that of another, (ii) the inherent kinetic properties of individual gSULTs, (iii) the relative availability of PAPS and sulfate-acceptor substrates, (iv) tissue and cell expression patterns of gPAPP and the various gSULTs, and (v) possible compensation by other phosphatases. "<-So even if Lithium inhibits gPAPP which may have height reducing effects if there's compensation by other phosphatases and/or the height beneficial effects of Lithium are greater than gPAPP inhibition then Lithium may lead to a net increase in height.


Lithium protects cartilage from cytokine-mediated degradation by reducing collagen-degrading MMP production via inhibition of the P38 mitogen-activated protein kinase pathway.

"To determine the effects and mechanism of action of lithium chloride (LiCl) on cartilage destruction induced by the pro-inflammatory cytokines IL-1, IL-1 + oncostatin M and TNF-α.
The release of collagen was assessed in bovine cartilage explant cultures, whereas collagenolytic activities (active and total) in conditioned culture supernatants were determined by bioassay.
LiCl, but not selective glycogen synthase kinase 3 (GSK-3) inhibitor compounds SB-415286 and TDZD-8, significantly decreased pro-inflammatory cytokine-induced collagen release from bovine cartilage via the down-regulation of collagenolytic activity. MMP-1 and MMP-13 expression was reduced in both bovine and human chondrocytes. LiCl selectively inhibited activation of the p38 mitogen-activated protein kinase pathway; effects that were recapitulated by specific p38 pathway inhibition.
LiCl can protect against cartilage damage induced by pro-inflammatory cytokines, and LiCl-mediated cartilage protection is not via a GSK-3-dependent mechanism, but potentially via inhibition of the p38 pathway."

Remember that inflammatory particles like TNF-alpha can reduce height by causing chondrocyte apoptosis.  So lithium can increase height by inhibiting inflammatory particles like TNF-alpha.

"we found that LiCl blocked cartilage collagen degradation in a bovine model and reduced levels of MMPs including the collagenases MMP-1 and MMP-13"

"LiCl reduced the level of activation of the p38 pathway substrate HSP27"<-It's speculated that Lithium Chloride stops IL-1 inflammatory cytokine collagen release through the p38 MAPK pathway.

"HSP27 was shown to be required for the IL-1 induction of the inflammatory mediators cyclooxygenase-2 and IL-6, at least in part, by the regulation of the stability of their mRNAs "

Phosphorylation of GSK-3beta by cGMP-dependent protein kinase II promotes hypertrophic differentiation of murine chondrocytes.[Lithium phosphorylates GSK-3Beta]

"cGMP-dependent protein kinase II (cGKII; encoded by PRKG2) is a serine/threonine kinase that is critical for skeletal growth in mammals; in mice, cGKII deficiency results in dwarfism. This growth defect [is] a consequence of an elongated growth plate and impaired chondrocyte hypertrophy. glycogen synthase kinase-3beta (GSK-3beta; encoded by Gsk3b) [is] a principal phosphorylation target of cGKII. In cultured mouse chondrocytes, phosphorylation-mediated inhibition of GSK-3beta was associated with enhanced hypertrophic differentiation{cGKII inhibits GSK-3beta which results in enhanced hypertrophic differentiation i.e. enhanced height growth}. cGKII induction of chondrocyte hypertrophy was suppressed by cotransfection with a phosphorylation-deficient mutant of GSK-3beta. Analyses of mice with compound deficiencies in both protein kinases (Prkg2(-/-)Gsk3b(+/-)) demonstrated that the growth retardation and elongated growth plate associated with cGKII deficiency were partially rescued by haploinsufficiency of Gsk3b. Beta-catenin levels decreased in Prkg2(-/-) mice, while overexpression of cGKII increased the accumulation and transactivation function of beta-catenin in mouse chondroprogenitor ATDC5 cells. This effect was blocked by coexpression of phosphorylation-deficient GSK-3beta. hypertrophic differentiation of growth plate chondrocytes during skeletal growth is promoted by phosphorylation and inactivation of GSK-3beta by cGKII."

Remember that cGKII has been found to increase height growth and is closely related to CNPs and Nitric Oxide.  Lithium inhibits GSK-3beta just like cGKII does, could Lithium have similar benefits on height growth?


Lithium treatment and thyroid abnormalities.

"The main findings from this review included: a) lithium definitely affects thyroid function as repeatedly shown by studies on cell cultures, experimental animals, volunteers, and patients; b) inhibition of thyroid hormone release is the critical mechanism in the development of hypothyroidism, goitre, and, perhaps, changes in the texture of the gland which are detected by ultrasonic scanning; c) compensatory mechanisms operate and prevent the development of hypothyroidism in the majority of patients; d) when additional risk factors are present, either environmental (such as iodine deficiency) or intrinsic (immunogenetic background), compensatory potential may be reduced and clinically relevant consequences may derive; e) hypothyroidism may develop in particular during the first years of lithium treatment, in middle-aged women, and in the presence of thyroid autoimmunity; f) thyroid autoimmunity is found in excess among patients suffering from affective disorders, irrespective of lithium exposure; g) in patients who have been on lithium for several years, the outcome of hypothyroidism, goitre, and thyroid autoimmunity do not much differ from those observed in the general population; h) hyperthyroidism and thyroid cancer are observed rarely during lithium treatment."

Lithium seems to have similar effects to thyroid hormone which is probably why supplementing with Lithium inhibits thyroid hormone production.

Lithium stimulates proliferation in cultured thyrocytes by activating Wnt/beta-catenin signalling.

"Lithium is well known to induce thyroid growth.  Thyroid proliferation depends on the thyroid-stimulating hormone (TSH)/cAMP/cAMP response element binding protein (CREB) pathway, [and] Wnt/beta-catenin signalling may be of critical importance{Wnt and beta-catenin are critical for bone growth as well}. In other cell types lithium activates canonical Wnt signalling by GSK-3beta inhibition, which in turn stabilizes cytosolic free beta-catenin. Here we investigated the potential modulation of Wnt/beta-catenin signalling under lithium treatment in primary and neoplastic human thyrocytes.
Primary (S18) and neoplastic (NPA, FTC133) thyrocytes treated with and without LiCl were analysed.
LiCl dose-dependently inhibited GSK-3beta, stabilized free beta-catenin and inhibited beta-catenin degradation. LiCl altered the assembly of adherens junction by upregulating the E-cad-herin repressor, Snail, and downregulated E-cadherin expression. At a dose of 5 mM, LiCl significantly increased the proliferative potency of thyrocytes, which appeared to be mediated by beta-catenin, since nuclear beta-catenin stimulated T-cell factor/lymphoid enhancer factor (TCF/LEF)-mediated transcription and upregulated downstream targets like cyclin D1. To characterize the specificity of Wnt/beta-catenin-driven thyrocyte proliferation, we transfected primary thyrocytes and FTC133 cells with dominant negative TCF4 to block Wnt-dependent pathways or with dominant negative CREB to inhibit the TSH/cAMP cascade. In cells transfected with dominant negative CREB lithium-stimulated proliferation was unchanged whereas blocking Wnt/beta-catenin by dominant negative TCF4 reduced proliferation by approx. 50%.
Wnt/beta-catenin signalling is of major importance in the control of lithium-dependent thyrocyte proliferation."

Lithium serves similar functions as Thryoid hormone.

Lithium is a GSK-3beta inhibitor which it accomplishes by adding a phosphate group to it. Other things that have been found to increases height like cGK II also operate by inhibiting GSK-3beta. Lithium in turn inhibits TNF-alpha which can cause chondrocyte apoptosis. However, Lithium inhibits gPAPP which is critical for height growth due to it adding phosphate groups to GAGs like chondroitin during key stages of endochondral ossification.  Lithium but not too much should increase height during development.

According to [Calcitonin and parathyroid hormone secretion and calcium metabolism in patients with diffuse toxic goiter during treatment with lithium carbonate], lithium induces an increase in interstitial calcium absorption.  According to Influence of lithium and exercise on serum levels of copper and zinc in rats., exercise decreases lithium concentration and lithium increases serum levels of zinc and copper.

Erythrocyte sodium-lithium countertransport activity is inversely correlated to adiponectin, retinol binding protein 4 and body height.

"Sodium/lithium countertransport (SLC) in the erythrocyte cell membrane is closely linked to obesity and insulin resistance. Adiponectin and retinol-binding protein 4 (RBP-4) are believed to affect obesity and insulin resistance. In the present study, we aimed to further characterize the relationship between SLC, inflammatory markers, adiponectin and RBP-4.
We included 93 clinically healthy 58-year-old men selected to display variations in insulin sensitivity. High sensitivity C-reactive protein (hs-CRP), TNF-alpha, soluble TNF-alpha-receptors (sTNFR) 1 and 2, IL-6 and RBP-4 were measured using antibody-based techniques. Adiponectin was determined by a radioimmunoassay kit. The lithium concentration in the special flux medium was measured by atomic absorption spectrophotometry.
In univariate analyses, SLC correlated negatively with RBP-4 (r(s) = -0.256, p = -0.017) and to adiponectin (r(s) = -0.316, p = 0.003) and positively with TNF-alpha (r(s) = 0.346, p = 0.001) and hs-CRP (r(s) = 0.288, p = 0.005). There were no statistically significant correlations with sTNFR 1 or 2 or IL-6. SLC was negatively associated to body height[Sodium Lithium countertransport decreases height?  What does that mean for lithium?} (r(s) = -0.256, p = 0.013).
Adiponectin is anti-inflammatory and anti-diabetic whereas RBP-4 supposedly decreases insulin sensitivity. The positive association of SLC with markers of inflammatory activity such as TNF-alpha and hs-CRP is in line with the proposed link between inflammation and insulin resistance."

"In human red cells, lithium is extruded against its own concentration gradient if the external medium contains sodium as a dominant cation in a process that does not require cellular ATP"<-This SLC.  If SLC is bad how do we affect this?  Do we want Lithium and Sodium levels low or high?

"insulin resistance could be the motor of the increased SLC activity"<-But insulin resistance increases height. Maybe SLC is only correlated with loss of height and not the cause.

"RBP-4 is a 21 kDa protein that binds and transports retinol and is mainly produced in adipose tissue and in the liver."

The study Sodium-Lithium Countertransport Has Low Affinity for Sodium in Hyperinsulinemic Hypertensive Subjects provides insight on how sodium & lithium concentrations affect SLC transport levels.

SLC peaked at around 100 Mm of Sodium for normatensive individuals and 150 Mm for hypertensive individuals. SLC was minimal at around 0 Mm for both. Eat the least amount of salt possible to grow taller. Insulin only affects SLC in hypertensive individuals.

The study did not mention any affects of Lithium levels on SLC.

Wnt/beta-catenin signaling stimulates chondrogenic and inhibits adipogenic differentiation of pericytes: potential relevance to vascular disease?

"Wnt/beta-catenin signaling regulates cell differentiation during embryonic and postnatal development, and there is increasing evidence that it is involved in vascular pathology. Wnt/beta-catenin signaling regulates the chondrogenic and adipogenic differentiation of pericytes[A nerve cell]. We demonstrate that pericytes express several Wnt receptors, including LDL receptor-related proteins 5 and 6, and Frizzled 1 to 4 and 7, 8, and 10, and that Wnt/beta-catenin signaling is stimulated by both Wnt3a and LiCl. Furthermore, induction of Wnt/beta-catenin signaling by LiCl enhances chondrogenesis in pericyte pellet cultures in the presence of transforming growth factor-beta3[LSJL stimulates the WNT pathway], as demonstrated by increased Sox-9 expression and glycosaminoglycan accumulation into the matrix. In contrast, transduction of pericytes with a recombinant adenovirus encoding dominant-negative T-cell factor-4 (RAd/dnTCF), which blocks Wnt/beta-catenin signaling, inhibited chondrogenesis, leading to reduced Sox-9 and type II collagen expression and less glycosaminoglycan accumulation."

"collagen II expression was not induced further by Wnt signaling, suggesting that its expression was already maximal."<-Maybe there's a maximal level for several genes after which height cannot be increased further by more of that gene?

The effect of lithium carbonate administration on growth in the domestic fowl.

"determine the effect of lithium carbonate administration on growth in a domestic fowl model. The results showed that lithium treatment at a therapeutic level (0.2-2.0 mEq/L) did not significantly alter plasma pH levels, food intake, fecal output or body weight gain. Bivariate and multivariate analysis of tibial dimensions revealed that lithium treatment primarily caused increased growth of proximal epiphyseal height. Allometric analyses indicated that lithium administration increased the growth differential between proximal and distal epiphyses."

Hypothalamic-pituitary-thyroid system activity during lithium augmentation therapy in patients with unipolar major depression.

"In 24 euthyroid patients with a major depressive episode who had not responded to antidepressant monotherapy of at least 4 weeks, we measured serum thyroid-stimulating hormone (TSH), total triiodothyronine (T3) and total thyroxine (T4) before (baseline) and during lithium augmentation therapy (follow-up). The time point of the endocrinological follow-up depended on the status of response, which was assessed weekly ). Responders were reassessed immediately after response was determined, and non-responders after 4 weeks of lithium augmentation.

There was a statistically significant change in thyroid system activity during lithium augmentation, with an increase of TSH levels and a decrease of peripheral T3 and T4 levels. However, there were no differences in any of the HPT hormones between responders and non-responders at baseline or at follow-up."

High Throughput Transcriptome Profiling of Lithium Stimulated Human Mesenchymal Stem Cells Reveals Priming towards Osteoblastic Lineage.

"Our results show suppression of proliferation and enhancement of alkaline phosphatase (ALP) activity upon lithium treatment of hMSCs under non-osteogenic conditions. Microarray profiling of lithium-stimulated hMSC revealed decreased expression of adipogenic genes (CEBPA, CMKLR1, HSD11B1) and genes involved in lipid biosynthesis. Interestingly, osteoclastogenic factors and immune responsive genes (IL7, IL8, CXCL1{up}, CXCL12, CCL20) were also downregulated. Negative transcriptional regulators of the osteogenic program (TWIST1 and PBX1) were suppressed while genes involved in mineralization like CLEC3B and ATF4 were induced. Gene ontology analysis revealed enrichment of upregulated genes related to mesenchymal cell differentiation and signal transduction. Lithium priming led to enhanced collagen 1 synthesis and osteogenic induction of lithium pretreated MSCs resulted in enhanced expression of Runx2, ALP and bone sialoprotein. However, siRNA-mediated knockdown of RRAD, CLEC3B and ATF4 attenuated lithium-induced osteogenic priming, identifying a role for RRAD, a member of small GTP binding protein family, in osteoblast differentiation."

This is contrary to evidence that Lithium increases MSC proliferation.

"LiCl at 5 mM resulted in more than 20% reduction while at 40 mM it had maximum inhibitory effect with more than 80% reduction in cell proliferation."

"lithium stimulation of hMSC for 7 days in the presence of expansion and osteogenic medium demonstrated highly inhibitory effect of 20 mM LiCl as no ALP staining was observed. However, lower concentration (5 mM) resulted in enhanced ALP staining under both culture conditions."

(Comparison using accession number and not just gene name to be done)
Genes upregulated by Lithium also upregulated by LSJL:
Edn1
A1BG
ALDOB{down}
KIRREL3
FLNC

Downregulated:
PTGS2{up}
CXCL1{up}
Kynu{up}
AREG{up}
RBMS1
NR4A2{up}
GPR115{up}
NR4A1{up}
CFI{up}
RASD1{up}
PACSIN1
COL12A1{up}
LAMA4{up}
WWTR1{up}
ODF4
PPFIBP1

%of Differentially regulated genes: 61.9%

Chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells treated by GSK-3 inhibitors.

"The present study has attempted to evaluate the effects of two well-known glycogen synthase kinase-3 inhibitors, including lithium chloride (LiCl) and SB216763 on a human marrow-derived MSC (hMSC) chondrogenic culture. Passaged-3 MSCs were condensed into small pellets and cultivated in the following groups based on the supplementation of chondrogenic medium: transforming growth factor (TGF)-β1, TGF-β1 + LiCl, TGF-β1 + SB216763, TGF-β3, TGF-β3 + LiCl, and TGF-β3 + SB216763. The cultures were maintained for 21 days and then analyzed for expression of Sox9, aggrecan, collagen II, β-catenin, and axin genes. Deposition of glycosaminoglycan (GAG) in the cartilage matrix was also measured for certain cultures. The presence of both LiCl and SB216763 along with TGF-β in the MSC chondrogenic culture led to the up-regulation of cartilage-specific genes. TGF-β3 appeared much better than TGF-β1. Based on our findings, SB216763 was more effective in up-regulation of cartilage-specific genes. These chondrogenic effects appeared to be mediated through the Wnt signaling pathway since β-catenin and axin tended to be up-regulated and down-regulated, respectively. In the culture with SB216763 + TGF-β3, significantly more GAG was deposited . Addition of either SB216763 or LiCl to hMSC chondrogenic culture up-regulates cartilage-specific gene expression and enhances GAG deposition in the culture."

"inhibition of GSK-3β was responsible for the therapeutic usefulness of lithium salts"

" the pellet cultures treated with LiCl appeared to have zones of hypertrophic chondrocytes "<-So Lithium encouraged more endochondral ossification.

The effects of Lithium on chondrogenic differentiation were not analyzed independently of the TGF-Beta growth factors.

Changes in body weight and body mass index among psychiatric patients receiving lithium, valproate, or topiramate: An open-label, nonrandomized chart review

Data for height was collected but not shown which is unfortunate.

Lithium, Bone and Body Weight Studies in Long-Term Lithium-Treated Patients and in the Rat

"Morphometric studies were carried out on radiographs taken on separate occasions, 2 years apart, from patients receiving long-term prophylactic lithium. Similar studies were made of femora of growing rats over a 1-year period. We conclude that lithium dos not affect the size or mineralization of bone from either humans or growing or growing rats, but that it affects whole body growth in growing rats."

Lithium treatment decreased median cortical width of the medicarpal.

There is no trend in any direction of length versus bone length in lithium versus control rates but that means it doesn't affect growth rate and not the length of growth so final growth could be higher.

Grow Taller with Ions

The Sodium-Potassium pump is an enzyme located in the plasma membrane of all cells including chondrocyte cells.  The maximum size achieved by chondrocytes in the hypertrophic zone during normal growth is one of the primary determinants of natural height growth.  If the cells grow too large then they burst.  Hypertrophic chondrocytes die but it is due to lack of nutrients not due to bursting.

Ions like Sodium, Potassium, and Lithium are charged so this may be one mechanism that Pulsed-Electric Magnetic Fields can influence height growth.  If you look at some of the studies listed there, you can see that 20-60% higher cell densities were achieved during the cell expanding phase.  The possible mechanism could be as mentioned earlier due to PEMF stimulating charged ions.

LSJL may help transport these ions into the chondrocyte cells by stimulating fluid flow and increasing pressure.   There's an LSJL study down below.

A key role for membrane transporter NKCC1 in mediating chondrocyte volume increase in the mammalian growth plate.

"The mechanisms that underlie growth plate chondrocyte volume increase and hence bone lengthening are poorly understood. Many cell types activate the Na-K-Cl cotransporter (NKCC) to bring about volume increase. We hypothesised that NKCC may be responsible for the volume expansion of hypertrophic chondrocytes. Metatarsals/metacarpals from 16 rat pups (P(7)) were incubated in the presence/absence of the specific NKCC inhibitor bumetanide and measurement of whole-bone lengths and histologic analysis of the growth plate were done after 24 hours. Fluorescent NKCC immunohistochemistry was visualised using a confocal laser scanning microscopy on seven rat tibial growth plates (P(7)). Microarray analysis was performed on mRNA isolated from proliferative and hypertrophic zone cells of tibial growth plates from five rats of each of three ages (P(49/53/58)). Exposure to bumetanide resulted in approximately 35% reduction of bone growth in a dose-dependent manner; histologic analysis showed that a reduction in hypertrophic zone height was responsible. Quantification of fluorescence immunohistochemistry revealed a significant (paired Student's t test, p < .05) change in NKCC from the intracellular space of proliferative cells to the cytosolic membrane of hypertrophic zone cells. Further, microarray analysis illustrated an increase in NKCC1 mRNA between proliferative and hypertrophic cells. The increase in NKCC1 mRNA in hypertrophic zone cells, its cellular localization, and reduced bone growth in the presence of the NKCC inhibitor bumetanide implicate NKCC in growth plate hypertrophic chondrocyte volume increase. Further investigation is warranted to determine the regulatory control of NKCC in the mammalian growth plate and the possible detrimental effect on bone growth with chronic exposure to loop diuretics."

The fact that there was only a 35% reduction in bone growth shows that there are other factors(like number of stem cells) but that's still a huge amount. 

The study states that the volume increase in hypertrophic chondrocytes occurs predominantly in the longitudinal direction which is why bones grow longer rather than wider in equal proportion.  "The volume of a typical cell increases from approximately 1000 µm3 in the [proliferative zone] to approximately 15,000 µm3 in the [hypertrophic zone]."  So after a certain point increasing the maximal size attained in chondrocyte hypertrophy while affect height a lot more than increasing the number of cells.

"[A change in the osmotic gradient] can result from a reduction in extracellular osmolarity, an increase in intracellular osmolarity, or a combination of the two."<-we try to change the extracellular osmolarity with hydrostatic pressure via LSJL.  Altering intracellular calcium secretions may alter intracellular osmolarity.

"That for cell swelling to be entirely mediated by a reduction in extracellular osmolarity, these cells would need to be exposed to an approximately 280 mOsmol reduction in osmolarity to approximately 120 mOsmol. Plasma osmolarity therefore would need to be reduced from approximately 300 mOsmol to near 20 mOsmol, which is clearly implausible. Therefore, osmotic gradient–driven water accumulation is more likely to be due to an increase in intracellular osmolarity in the transition of a PZC to an HZC."<-This has been confirmed by the fact that a huge amount of hydrostatic pressure is required to induce chondrogenesis and LSJL only induces a small amount of hydrostatic pressure however LSJL may alter intracellular osmolarity thereby altering intracellular calcium secretions.

"Intracellular osmolarity can be raised by the catabolism of intracellular macromolecules (eg, proteins and/or complex carbohydrates) into osmotically active components (eg, amino acids and/or simple sugars)... The accumulation of organic solutes (eg, simple sugars and amino acids) is responsible for approximately 9% of the osmolytes required"

"The putative presence of transporters other than NKCC1 and/or redundancy in the system to drive HZC volume increase may explain the reduced size of NKCC1 knockout mice over the first 2 weeks, a period of maximal growth rate, but their ability to recover to the size of wild-type animals as they reach maturity.  This ability of NKCC1 knockouts to recover may explain why there are limited reports of skeletal growth retardation in children exposed to loop diuretics (eg, bumetanide and furosemide). "

Chloride might be the key chondrocyte stimulating compound. 

Chloride channels regulate chondrogenesis in chicken mandibular mesenchymal cells.  

"Voltage gated chloride channels (ClCs) play an important role in the regulation of intracellular pH and cell volume homeostasis[So any insights into chloride channels can provide insights into how intracellular pH and cell volume increases height]. Mutations of these genes result in genetic diseases with abnormal bone deformation and body size, indicating that ClCs may have a role in chondrogenesis. In the present study, we isolated chicken mandibular mesenchymal cells (CMMC) from Hamburg-Hamilton (HH) stage 26 chick embryos and induced chondrocyte maturation by using ascorbic acid[Vitamin C] and β-glycerophosphate (AA-BGP). We also determined the effect of the chloride channel inhibitor NPPB [5-nitro-2-(3-phenylpropylamino) benzoic acid] on regulation of growth, differentiation, and gene expression in these cells using MTT and real-time PCR assays. We found that CLCN1 and CLCN3-7 mRNA were expressed in CMMC and NPPB reduced expression of CLCN3{down in LSJL}, CLCN5, and CLCN7 mRNA in these cells. At the same time, NPPB inhibited the growth of the CMMC, but had no effect on the mRNA level of cyclin D1 and cyclin E (P>0.05) with/without AA-BGP treatment. AA-BGP increased markers for early chondrocyte differentiation including type II collagen, aggrecan (P<0.01) and Sox9 (P<0.05), whilst had no effect on the late chondrocyte differentiation marker type X collagen. NPPB antagonized AA-BGP-induced expression of type II collagen and aggrecan (P<0.05). Furthermore, NPPB downregulated type X collagen (P<0.05) with/without AA-BGP treatment. We conclude that abundant chloride channel genes in CMMC play important roles in regulating chondrocyte proliferation and differentiation. Type X collagen might function as a target of chloride channel inhibitors during the differentiation process[Chloride Channel inhibitors downregulated Type X Collagen which is a terminal differentiation marker but since the Chloride Channel inhibitor decreased the growth of the Mesenchymal Cells maybe Type X Collagen is essential for maximizing chondrocyte growth]."

"During embryonic development, chondrogenesis begins with mesenchymal cell recruitment and migration, proliferation, and condensation."<-This is what we try to achieve with LSJL.  Hydrostatic Pressure to recruit  MSCs to the site of the pressure so that the new cartilage acts as a sponge for the pressure.  Then that cartilage undergoes endochondral ossification within the bone and makes a person taller.

"ClC is associated with the volume-activated chloride current and is involved in cell volume regulation"

"endogenous ClC-3 is associated with the volume-activated chloride current and is involved in cell volume regulation. In our study, NPPB suppressed the expression of ClC-3, which may influence chondrocyte swelling (hypertrophy) and further influence the synthesis of type X collagen."<-maximizing the potential of the volume-activated chloride current will help height growth and further evidence that Type X Collagen should not be inhibited but may in fact help maximize growth.

The osmotic sensitivity of rat growth plate chondrocytes in situ; clarifying the mechanisms of hypertrophy. 

"Bone elongation is predominantly driven by the volume expansion of growth plate chondrocytes. This mechanism was initially believed to be "hypertrophy", describing a proportional increase of cell water and organelles. However, morphometrical analysis subsequently assumed the increase to be "swelling", resulting in a disproportionate increase of cell water (osmotically active fraction). Histological approaches were performed on fixed tissue, and for the "swelling" assumption to be valid, the osmotic sensitivity of living cells before and during volume increase should differ. To test this, analysis of images acquired by 2-photon laser scanning microscopy (2PLSM) were used to determine the osmotic sensitivity, and osmotically active/inactive proportions of in situ chondrocytes from 15 living rat growth plates exposed to varying media osmolarities ( approximately 0-580 mOsm). The dimensions of cell volume swelling in hypotonic[hypotonic means that there was less water outside the cell then in cause water to be taken into the cell making it grow larger] media were different to the preferential lengthening seen in vivo, confirming the complexity of directional cell volume increase. Boyle-van't Hoff analysis of cell volume over the range of media osmolarity indicated no significant difference (Student's t-test) in the osmotically inactive fraction, 39.5 +/- 2.9% and 47.0 +/- 4.3% (n = 13) for proliferative and hypertrophic zones, respectively, or the sensitivity of volume to changes in media osmolarity (proliferative 15.5 +/- 0.8 and hypertrophic zone 15.5 +/- 1.2%volume . Osm). The osmotic fractions did not change as chondrocytes progress from proliferative to hypertrophic regions of the growth plate. Our data suggest cell volume increase by hypertrophy may play a greater role in cell enlargement than swelling, and should be re-evaluated as a mechanism responsible for growth plate chondrocyte volume increase and hence bone elongation." 

This suggests that water may not be the primary force in chondrocyte hypertrophy.

"More recently, however, stereological analysis of fixed tissue has shown a disproportionate increase in cell compartments. These observations showed an increase in cell organelle volumes (endoplasmic reticulum, Golgi membranes, and mitochondria) was responsible for ∼15% of the total cell volume increase between proliferative zone chondrocytes (PZC) and hypertrophic zone chondrocytes (HZC), the majority (85%) of volume increase arising from cytoplasm and nucleoplasm expansion"

"The apparent preferential increase of cell cytoplasm and nucleoplasm led to the assumption that cell volume increase was primarily due to “swelling” (fluid accumulation)."<-so swelling is still a part of the cell volume increase

" The osmotically active fraction describes freely mobile water within the cell, and it is this fraction that would increase dramatically if a cell increased in volume by “swelling” alone. Conversely, if a cell showed a general increase in total volume in proportion to its intracellular constituents, “hypertrophy,” then the osmotically active and inactive fractions will increase in proportion. If, as stereological approaches have suggested, cell volume increase occurs primarily due to fluid accumulation rather than hypertrophy, the osmotically inactive/active fractions of the PZC and HZC should differ"<-If much of the size increase is due to hypertrophy that doesn't mean that some is due to swelling.

"Cell volumes increased ∼10-fold, from 1,172 ± 209µm3 in S1 to 10,584 ± 1,315 µm3 in S8"<-this is a pretty large change and illustrates the importance of chondrocyte hypertrophy.

"The increase in volume shown by this PZC was from 768 µm³ in 280 mOsm media to 1,900 µm³ in near 0 mOsm, whereas the HZC volume increased from 6,236 µm³ in 280 mOsm media to 26,878 µm³ in near 0 mOsm media."<-Higher osmolarity meaning a higher concentration of a substance other than water so less hydrostatic pressure. More water still increased swelling it was just the same in just proliferative and hypertrophic zones.

"All cells showed a clear reduction in volume with hyper-osmotic challenge, and hypotonicity increased chondrocyte volume even when media osmolarity reduced from 80 to ∼0 mOsm media, with little apparent restriction to swelling from the surrounding matrix"<-the higher the water concentration(or the lower the concentration of other solutes) the higher the chondrocyte volume.

"GAGs will preferentially attract cations, but matrix/bone restriction to swelling will limit the volume of osmotically obliged water that can follow."<-So GAGs will absorb water.  Since the amount of GAGs increase based on layer this could be why the cells don't change in osmotic sensitivity as it is the level of GAGs that are changing.

"The concentration and the type of GAG have been shown to vary from proliferative to hypertrophic zones"<-So the osmolaric sensitivity varies with the GAGs

"We also cannot discount that the driving force for cell volume increase is by swelling, followed by consolidation through the production of “dry matter.” A PZC increasing in volume by 10-fold through “hypertrophy,” with a concomitant increase in osmotically inactive volume would require ∼3,600 µm³ (assuming a 40% osmotically inactive fraction) increase in “dry matter.” Transport of the components required for synthesis of osmotically inactive structures (e.g., amino acids, simple sugars, etc.) may cause a transient osmotic gradient inducing cell swelling. Indeed, transporter (with receptor) mRNA expression shows a near tripling in the number of relevant up-regulated genes"<-Water may still be involved indirectly.

LSJL may involve ionic transport.

Knee-loading modality drives molecular transport in mouse femur.

"Load-driven interstitial fluid flow and molecular transport[Calcium gate ion channels show the importance of both calcium and barium] play a role in the enhancement of bone formation. In order to evaluate load-driven molecular transport in a lacunocanalicular network, we conducted fluorescence recovery after photobleaching (FRAP) experiments using lacunae stained with uranine (376 Da). Loads were applied to a mouse femur ex vivo with a novel knee-loading modality, where the distal epiphysis was loaded with a sinusoidal force at 2 Hz[40V at 1.7N]. The lacunae in the diaphysis located 25% (approximately 4 mm) proximal to the loading site were photobleached and sequentially imaged, and a time constant for fluorescence recovery was determined both with and without knee loading. The time constant was estimated as the period to recover 63% of fluorescent intensity using a best-fit exponential curve. The results reveal that the applied loads shortened the time constant from 33 +/- 9 s with non-loading control to 25 +/- 11 s with knee loading. The strain in the measurement site was <100 microstain along the femoral midshaft, which was an order of magnitude smaller than the minimum effective strain threshold for bone remodeling. molecular transport in cortical bone is enhanced by the loads applied to the epiphysis."

"Dynamic deformation of porous bone matrix is considered to stimulate interstitial fluid flow and molecular transport."

"Transport of varying molecules such as ions, nutrients, and waste materials within lacunae is mainly governed by a process of diffusion and convection. Mechanical loads are considered to facilitate their transport through a network of canaliculi as well as Haversian and Volkmann’s canals"

"the loading force was estimated as 0.042 N per voltage to the loader. In this study sinusoidal loads at the loading frequency of 2 Hz with 1–4 N (peak-to-peak) force were employed."

"The observed enhancement of molecular transport in response to the applied loads indicates periodic load-driven fluid mixing through the canalicular network."

"The observed time constants of fluorescence recovery clearly showed enhanced molecular transport with knee loading"<-All molecules and that's a wide category, some of those molecules are beneficial for height increase.

"The described knee-loading modality may remotely displace fluid from the epiphysis towards the metaphysis and the diaphysis and stimulate molecular transport in cortical bone."<-this displacement increases hydrostatic pressure and is partly what induces chondrogenesis.

"the strain in the femur 25% (∼4 mm) proximal to the distal end of the femur was measured as 26.8 ± 7.8, 43 ± 7.7 and 93.8 ± 8.2 μstrain with 1, 2 and 4 N force, respectively. The best-fit regression line for the force–strain relationship was y = 22.7x + 1.5 with r 2 = 0.99. Note that the background strain level without any loads was ∼10 μstrain."<-So strain increases rapidly as force increases.  To get 1500 micorstain you'd need to use about 66N.  But this did not measure strain directly at the epiphysis.

But an increase in molecular and ionic transport could play a role in the effects of LSJL.

Switch of voltage-gated K+ channel expression in the plasma membrane of chondrogenic cells affects cytosolic Ca2+-oscillations and cartilage formation.

"Using patch-clamp, RT-PCR and Western-blot experiments, we found that chondrogenic cells in primary micromass cell cultures obtained from embryonic chicken limb buds expressed voltage-gated Na(V)1.4, K(V)1.1, K(V)1.3 and K(V)4.1 channels, although K(V)1.3 was not detectable in the plasma membrane. Tetrodotoxin (TTX), the inhibitor of Na(V)1.4 channels, had no effect on cartilage formation[so sodium likely doesn't affect chondrogenesis]. In contrast, presence of 20 mM of the K(+) channel blocker tetraethyl-ammonium (TEA) during the time-window of the final commitment of chondrogenic cells reduced K(V) currents (to 27±3% of control), cell proliferation (thymidine incorporation: to 39±4.4% of control), expression of cartilage-specific genes and consequently, cartilage formation (metachromasia: to 18.0±6.4% of control) and also depolarized the membrane potential (by 9.3±2.1 mV)[potassium plays a large role in chondrogenesis]. High-frequency Ca(2+)-oscillations were also suppressed by 10 mM TEA (confocal microscopy: frequency to 8.5±2.6% of the control). Peak expression of TEA-sensitive K(V)1.1 in the plasma membrane overlapped with this period. Application of TEA to differentiated chondrocytes, mainly expressing the TEA-insensitive K(V)4.1 did not affect cartilage formation.
These data demonstrate that the differentiation and proliferation of chondrogenic cells depend on rapid Ca(2+)-oscillations, which are modulated by K(V)-driven membrane potential changes. K(V)1.1 function seems especially critical during the final commitment period. We show the critical role of voltage-gated cation channels in the differentiation of non-excitable cells with potential therapeutic use."

So altering the K+ channels may be a way to grow taller.

"Differentiating chondrocytes in high density cultures (HDC) displayed rapid and repetitive [Ca2+]i transients"<-So inducing rapid and repetitive [Ca2+]i transients in epiphyseal MSCs may be a way to grow taller.

"With differentiation, the frequency of the transients decreased, (0.163±0.020 Hz on day 1 and 0.089±0.010 Hz on day 3, ANOVA: p<0.01, Holm-Sidak (HS): t=3.719) but their amplitude did not change significantly (expressed as F/F0: 1.98±0.40 on day 1 and 1.64±0.17 on day 3; ANOVA: p>0.05)"<-So the key is to increase the frequency of [Ca2+]i transients.

"The depolarization of the cell membrane by administration of 20 mM KCl on day 2 resulted in a large and transient increase in [Ca2+]i"

"Upon the removal of calcium from the external medium the amplitude and frequency of the repetitive calcium transients were decreased, sometimes completely eliminated"

"chondrogenic mesenchymal cells exhibited unique Ca2+ oscillations in our experiments"<-mimic the Ca2+ oscillations in mesenchymal cells to induce chondrogenesis.

"human mesenchymal stem cells [Ca2+]i oscillations [have] an average frequency of 1 transient in 120 sec were reported"

"cells of HDC exhibited spontaneous oscillations with an extraordinarily high frequency (approx. 5 in 60 sec on day 1)"<-high density culture induces chondrogenesis so our goal should be 5 oscillations per minute.

Frequency is important to LSJL with the optimal frequency being 0.5Hz this could be related to the calcium ion oscillations.

Suppression of mammalian bone growth by membrane transport inhibitors.

"We investigated the role of the Na(+) /H(+) antiporter (NHE1)[regulates pH and cell volume] and anion exchanger (AE2) in bone lengthening and GP chondrocyte hypertrophy in Sprague-Dawley 7-day-old rat (P7) bone rudiments using the inhibitors EIPA (5-(N-ethyl-N-isopropyl)amiloride) and DIDS (4,4-diidothiocyano-2,2-stilbenedisulphonate) respectively. We have also determined cell-associated levels of these transporters along the GP using fluorescent immunohistochemistry (FIHC). Culture of bones with EIPA or DIDS inhibited rudiment growth (50% at approx. 250µM and 25µM respectively). Both decreased the size of the hypertrophic zone (P<0.05) but had no effect on overall length or cell density of the GP. In situ chondrocyte volume in proliferative and hypertrophic zones was decreased (P<0.01) with EIPA but not DIDS. FIHC labelling of NHE1 was relatively high and constant along the GP but declined steeply in the late hypertrophic zone. In contrast, AE2 labelling was relatively low in proliferative zone cells but increased (P<0.05) reaching a maximum in the early hypertrophic zone, before falling rapidly in the late hypertrophic zone suggesting AE2 might regulate the transition phase of chondrocytes between proliferative and hypertrophic zones. The inhibition of bone growth by EIPA may be due to a reduction to chondrocyte volume set-point. However the effect of DIDS could result from inhibition of AE2 and blocking of the transition phase."

"The controlled increase in chondrocyte size and the regulation of the intracellular environment (e.g. ion content, pHi) to permit optimal metabolism must be closely regulated throughout the hypertrophic process mainly by the activity of the membrane transporters in the chondrocyte membrane. It is likely that there is an important role for transporters which regulate the movement of Na+ and anions (e.g. HCO3-) across the cell membrane"

"Increased activity of NHE1 combined with the parallel activation of the anion exchanger isoform AE2 occurs following the shrinking of cells in media of raised osmolarity. This gives rise to the recovery of cell volume through the process of regulatory volume increase (RVI) and also the maintenance of pHi in the face of the osmotic challenge"

"Activation of these transporters without a change in extracellular osmotic pressure, will directly increase cell volume and re-set pHi"<-so increasing NHE1 and AE2 will help you grow taller.

"EIPA significantly reduced the volume of chondrocytes throughout the length of the GP whereas DIDS had no significant effect"

"levels of NHE1 remained high in chondrocytes throughout most of the GP, but declined in late hypertrophic cells whereas AE2 levels demonstrated a biphasic effect"

"DIDS is a relatively non-specific inhibitor of AEs, as it also blocks Cl- channels in a range of cell types [whereas EIPA is a specific inhibitor of NHEs]"

"DIDS has also been reported to block diastrophic dysplasia sulphate transport (DTDST)-mediated SO4 - uptake which is essential for proteoglycan synthesis"

"programmed cell death (‘apoptosis’) in some cell types is associated with a decline in AE2

levels"

"the addition of  IGF-1 to isolated chondrocytes stimulates NHE1 potentially resulting in an optimal pHi for the early stages of chondrocyte hypertrophy."

Effect of sodium selenite on growth, insulin-like growth factor-binding proteins and insulin-like growth factor-I in rats.

"30 male Wistar rats were randomized into three groups. Group A was treated with sodium selenite in the drinking water (3.3 mg selenium/l). Group B was ad libitum fed with free access to standard fodder and tap water and group C was pair fed relative to the selenium-treated rats. Serum IGF-I and IGFBPs were determined on days 0, 14 and at the end of the study on day 35. Selenium-treated rats had significantly lower body weights compared with group B rats on day 9 and group C rats on day 14. Tibia length was measured at the end of the study and no difference was observed between groups B and C (3.77 +/- 0.04 cm vs 3.60 +/- 0.02 cm); however, selenium-treated rats had significantly shorter tibia lengths (3.46 +/- 0.03 cm) compared with rats in groups B and C. Selenium treatment induced a significant reduction in circulating IGF-I by the end of the study compared with ad libitum and pair fed rats. Serum subjected to Western ligand blots showed four distinct IGFBP bands with apparent relative molecular weights of 38-47 kDa (doublet) (IGFBP-3), 30 kDa (IGFBP-1 and/or IGFBP-2) and 24 kDa (IGFBP-4)."

Couldn't access full study.


Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus.

"ATP production was dose-dependently decreased by NaF(Sodium Fluoride) in the human chondrocytic cell line HCS-2/8. In addition, both chondrocyte proliferation and differentiation were inhibited, whereas cell death was promoted by treatment with NaF. Combinational treatment with NaF and lactate enhanced translocation of phospho-Smad1/5/8 to the nucleus, as well as gene expression of ALP, VEGF, COL10a1, and MMP13, which were the markers of late mature and hypertrophic chondrocytes. Furthermore, the production of type X collagen and activation of MMP9 were also promoted under the same conditions."

"decreased ATP production by NaF promotes hypertrophy-like changes via activation of phospho-Smad1/5/8 in the presence of lactate."

"cartilage is an avascular tissue and chondrocytes generate energy in the form of adenosine triphosphate (ATP), mostly dependent on anaerobic metabolism of glucose. Therefore, if this metabolism is impaired, intracellular ATP production is profoundly decreased; and the chondrocytes are forced to degenerate, following the pathway to hypertrophy-like changes."

"There was no recovery of the decreased intracellular ATP production when lactate was present"<- Lactate is an end product of glycolosis.

"the gene expression of MCT2 was remarkably increased in HCS-2/8 cells treated with NaF or the NaF combined with lactate in the comparison with that in the untreated ones"

"MCT1 protein was decreased in pellet culture of RCS cells treated with NaF, but MCT2 protein was increased"

"Sodium lactate enhances the BMP signaling but not nuclear factor (NF)-kB signaling under the condition of lowered ATP production in HCS-2/8 cells.  MCT1 contributes to cell death through activation of NF-kB in mouse chondrocytic ATDC5 cells"

"Gene expression of COL10a1, ALP, MMP13, and VEGF, all of which are markers of hypertrophic

chondrocytes, was promoted by treatment with NaF or it in combination with lactate more strongly than by that with PBS or lactate alone"

"the gene expression of AGGRECAN (ACAN), which is a marker of mature chondrocytes, was decreased in the cells treated with NaF or NaF combined with lactate compared with its expression in the presence of PBS or lactate alone."

"the active form of MMP9, which is another marker of hypertrophic chondrocytes, was detected in medium conditioned by HCS-2/8 cells treated with NaF or NaF combined with lactate"

Note that these scientists are looking at articular cartilage not growth plate cartilage where hypertrophy is beneficial.

"stimulation by lactate leads to activation of signal transducers and activators of transcription 3 (STAT3) and extracellular signal-regulated kinase (ERK)1/2 signaling pathways in human mesenchymal stem cells"

A chemical genetics strategy exposes novel modulators of chondrogenesis that act by blocking a potassium channel, Kcnd2, & reveals a potential role for potassium channels in limb development

"[We used] primary cultures of murine limb bud-derived mesenchymal (PLM) cells. Chondroblast differentiation is associated with increased SOX5, 6 and 9 activity; while hypertrophic differentiation is associated with reduced SOX5, 6 and 9 activity. Therefore, a SOX5/6/9-responsive reporter gene was used to follow expression of the chondroblast phenotype. Two compounds identified, Butamben (butyl 4-aminobenzoate; BAB) and Phenazopyridine hydrochloride (PHCl), exhibited strong pro-chondrogenic activity and morphologically similar alcian blue staining. BAB is a member of the benzocaine family of analgesics and functions by inhibiting sodium channel activity. However, BAB has also been shown to have potassium channel-blocking activity. Specifically, BAB inhibits the activity of Kcnd2; which through transcriptional profiling was also found to be down-regulated by bone morphogenetic protein-4 (BMP4). BAB and PHCl may be able to modulate chondrogenesis by acting on potassium channels. To confirm this idea we examined molecular activities of PLM cultures treated with BAB and PHCl at two stages of chondrogenesis: 1. pre-chondrocyte to chondroblast and 2. chondrocyte to hypertrophic chondrocyte. Results confirm, BAB and PHCl increase expression of chondrogenic markers and reduce expression of hypertrophic markers. In addition patch clamp analysis revealed both BAB and PHCl are able to block, at least partially, KCND2 channel activity."

30-45 Mm of BAB was optimal for chondroinduction.

"BAB (10 μM) was found to maximally inhibit [KCN2] channel activity by approximately 30%, and increasing the dose had no further impact on channel activity"

"PHCl is a more effective KCND2 channel blocker than BAB, and at higher concentrations PHCL exhibits > 80 % inhibition of KCND2 channel activity"

DIELECTRIC PROPERTIES OF CHONDROCYTES IN A CHANGING ENVIRONMENT.

"Chondrocytes, the only cell type that produces cartilage, occupy a unique bioelectrical environment. Mechanical load generates bioelectrical stimulation through rapid changes in ionic and water content around cells. A chondrocytes response to changing ionic gradients plays an essential role in regulation of extra cellular matrix synthesis. An abnormal response by the cell may result in abnormal deposition of proteins leading to degeneration of tissue with pathological consequences. Typically, response to ionic and osmotic gradients across cell membranes is by ion movement through channels as a cell reacts to maintain equilibrium. Chondrocytes possess an array of ion channels on par with electrically excitable tissues. Ion [channels are present in] chondrocytes. This proposal will quantify in real time dielectric properties of the membrane and cytoplasm in response to electrochemical changes to cells related to ion channel gene expression. We hypothesize that dielectric response of chondrocytes to changes in environment will be dependent upon ion channel activity. The objectives of this proposal are (1) to document dielectric changes in chondrocytes in real time when exposed to changes in osmolality, pH and ion flow using a novel microfluidic device. (2) Measure gene expression of ion channels in chondrocytes as a response to these changes. (3) Chondrocytes function under hypoxic conditions. We will directly compare dielectric response under normal versus hypoxic conditions and identify ion channels active in chondrocytes under hypoxic conditions. To our knowledge, our results will be the first description of dielectric properties of chondrocytes measured in real time and correlated to ion channel expression under hypoxia and will fundamentally impact cartilage biology unifying biomechanical and bioelectrical events in cartilage through ion channel response."

Roles of Sodium Hydrogen Exchanger (NHE1) and Anion Exchanger (AE2) across Chondrocytes Plasma Membrane during Longitudinal Bone Growth

"Mammalian long bone growth occurs through endochondral ossification, majorly regulated by the controlled enlargement of chondrocytes at the growth plate (GP). This study aimed to investigate the roles of Na+/H+ (sodium hydrogen exchanger (NHE1)) and HCO3- (anion exchanger [AE2]) during longitudinal bone growth in mammals. Bones from P10 Sprague Dawley rat pups were cultured ex vivo in the presence or absence of NHE1 and AE2 inhibitors to determine their effect on long bone growth. Gross morphometry, histomorphometry, and immunohistochemistry were used to assess the bone growth. The results revealed that the culture of the bones in the presence of NHE1 and AE2 inhibitors reduces bone growth significantly by approximately 11%. The inhibitor significantly  reduces bone growth velocity and the length of the hypertrophic chondrocyte zone without any effect on the total GP length. The total GP chondrocyte density was significantly reduced, but hypertrophic chondrocyte densities remained constant. NHE1 fluorescence signaling across the GP length was higher than AE2, and their localization was significantly  inhibited at the hypertrophic chondrocytes zone. The GP lengthening was majorly driven by an increase in the overall GP chondrocyte and hypertrophic chondrocyte densities apart from the regulatory volume phenomenon. This may suggest that NHE1 and AE2 could have a regulatory role in long bone growth."

"The resting zone contained stem-like chondroprogenitor cells that were believed to have a limited exhausted differentiation and proliferative capacity in all mammals except rodents. The finite proliferative capacity brings about growth plate fusion and the subsequent cessation of long bone growth, the cells in this zone are scattered within the cartilage matrix and usually have low rates of proliferation"

"plasma membrane transporters are implicated through the osmotic regulation of chondrocytes volume hypertrophy due to ion exchanges within the hypertrophic chondrocyte boundaries."

"mammalian hypertrophic chondrocytes undergo volume increase that subsequently contribute to long bone growth: (1) by a proportionate increase in dry mass production and fluid intake known as true hypertrophy, (2) by chondrocytes swelling, and (3) through a proportionate increase in dry mass along with fluid volume increase."

Even if there is a cell cycle limitation via senescence a change in chondrocyte hypertrophy will still increase height.

"NKCC1 plasma membrane transporter equally has a role in whole bone lengthening and growth plate hypertrophic chondrocyte volume regulation that positively increases bone growth."

Grow Taller with Acai Berry and Lithium?

A lot of times where the weight loss community goes, the height increase community follows.  One of the things hot in the weight loss community is Acai Berry.  Does Acai Berry have any possible usefulness for height gain?


Antioxidant, mutagenic, and antimutagenic activity of frozen fruits.


"Fresh fruits [may inhibit] reactive species and free radicals. 23 samples of frozen fruits were analyzed for their nutritional composition, total polyphenols, total carotenoids, and vitamin C content. Antioxidant, mutagenic, and antimutagenic effects were also evaluated. Antioxidant assays included 2,2-diphenyl-1-picrylhydrazyl radical (DPPH(.)) scavenging activity and determination of superoxide dismutase (SOD)- and catalase (CAT)-like activities. Mutagenic and antimutagenic evaluations were performed in eukaryotic cells of Saccharomyces cerevisiae yeast. Most samples (74%) showed antioxidant activity similar to vitamin C in the DPPH(.) assay, and this activity was positively correlated (r = 0.366; P Only four fruits (acai, cashew apple, kiwi fruit, and strawberry) showed mutagenic activity when tested in high (5%, 10%, and 15% [wt/vol]) concentrations. Twelve samples presented antimutagenic effects against hydrogen peroxide, and this effect was positively correlated with CAT-like activity (r = 0.400; P)."

Mutagenic refers to a substances ability to cause a mutation in the genetic code.  Obviously this effect is very powerful but we need to cause a specific mutation in order to influence height.  The ability of acai berry to fight free radicals can have a positive effect on free radicals too as it is a possibility that free radicals might negatively affect chondrocytes in the growth plate.


Antioxidant capacity and other bioactivities of the freeze-dried Amazonian palm berry, Euterpe oleraceae mart. (acai).


"The fruit of Euterpe oleraceae, commonly known as acai, has been demonstrated to exhibit significantly high antioxidant capacity in vitro, especially for superoxide and peroxyl scavenging, and, therefore, may have possible health benefits. The antioxidant capacities of freeze-dried acai fruit pulp/skin powder (OptiAcai) were evaluated by different assays with various free radical sources. [Acai has] exceptional activity against superoxide in the superoxide scavenging (SOD) assay, the highest of any food reported to date against the peroxyl radical as measured by the oxygen radical absorbance capacity assay, and mild activity against both the peroxynitrite and hydroxyl radical by the peroxynitrite averting capacity (NORAC) and hydroxyl radical averting capacity (HORAC) assays, respectively. The SOD of acai was 1614 units/g, an extremely high scavenging capacity for O2*-, by far the highest of any fruit or vegetable tested to date. Total phenolics were also tested as comparison. In the total antioxidant (TAO) assay, antioxidants in acai were differentiated into "slow-acting" and "fast-acting" components. An assay measuring inhibition of reactive oxygen species (ROS) formation in freshly purified human neutrophils showed that antioxidants in acai are able to enter human cells in a fully functional form and to perform an oxygen quenching function at very low doses.  Acai was found to be a potential cyclooxygenase (COX)-1 and COX-2 inhibitor [and] showed a weak effect on lipopolysaccharide (LPS)-induced nitric oxide but no effect on either lymphocyte proliferation and phagocytic capacity."


Acai berry inhibits Cox2 and Cox2 affects height growth. Cox2 is biphasic so there is an equilibrium quantity involved. Fighting free radicals would help if the levels of free radicals are above an equilibrium threshold.

Conclusin, Acai Berry is one of the best sources of anti-oxidants with which to fight free radicals. Acai berry has mutagenic effects but these mutagenic effects may not increase height.  One mutagenic substance that may increase height could be Lithium:


Review of lithium effects on brain and blood.


"Lithium acts through multiple pathways to inhibit glycogen synthetase kinase-3beta (GSK3 beta). This enzyme phosphorylates and inhibits nuclear factors that turn on cell growth[including possibly growth plate cells] and protection programs, including the nuclear factor of activated T cells (NFAT) and WNT/beta-catenin. In animals, lithium upregulates neurotrophins, including brain-derived neurotrophic factor (BDNF), nerve growth factor, neurotrophin-3 (NT3), as well as receptors to these growth factors in brain. Lithium also stimulates proliferation of stem cells, including bone marrow and neural stem cells in the subventricular zone, striatum, and forebrain. The stimulation of endogenous neural stem cells may explain why lithium increases brain cell density and volume in patients with bipolar disorders. Lithium also increases brain concentrations of the neuronal markers n-acetyl-aspartate and myoinositol. Lithium also remarkably protects neurons against glutamate, seizures, and apoptosis due to a wide variety of neurotoxins. The effective dose range for lithium is 0.6-1.0 mM in serum and >1.5 mM may be toxic. Serum lithium levels of 1.5-2.0 mM may have mild and reversible toxic effects on kidney, liver, heart, and glands. Serum levels of >2 mM may be associated with neurological symptoms, including cerebellar dysfunction. Prolonged lithium intoxication >2 mM can cause permanent brain damage. Lithium has low mutagenic and carcinogenic risk."

Lithium is not available in foods as far as I know so it may be valuable if you happen to have a prescription for it.