Grow Taller with Folinic Acid

For the first time, there is evidence that Folinic Acid treatment may increase length in developing individuals see (*NEW*).  Considering that Folinic Acid should not be harmful to length development, if an individual has active growth plates it's definitely a supplement one should add to their regime.

Hypermethylation of the growth plate chondrocytes may be a way to induce supranatural height growth.  DNA Methyltransferase stopping the addition of methyl groups to stem cell chondrocytes in the resting zone may be an indication to cease height growth.  High levels of HGH may also cause hypermethylation explaining gigantism(but why Gigantism is not inducible in all with HGH is unclear).  Folinic Acid is available as a supplement:Folinic Acid 800 mcg.

Are there any other ways of inducing DNA Hypermethylation(Bare in mind that DNA Hypermethylation increases the spread of cancer as DNA Methylation is one of the negative feedback mechanisms on cell growth)?  And since hypermethylation may not be possible to induce in growth plates general methods of increasing DNA synthesis like Folinic Acid may have similar effects.

Damaging effects of chronic low-dose methotrexate usage on primary bone formation in young rats and potential protective effects of folinic acid supplementary treatment.

"Methotrexate (MTX) is a most commonly used anti-metabolite in cancer treatment and as an anti-rheumatic drug. MTX chemotherapy at a high dose is known to cause bone growth defects in growing bones. We examined effects on bone growth of long-term MTX chemotherapy at a low dose in young rats, and potential protective effects of supplementary treatment with antidote folinic acid (given ip at 1 mg/kg 6 h after MTX). After two cycles of 5 once-daily MTX injections (at 0.75 mg/kg, 5 days on/9 days off/5 days on), MTX at this dose caused significant reduction in heights of growth plate and primary spongiosa bone on day 22 compared to controls. In contrast, a similar dosing regimen but at a lower dose (0.4 mg/kg) caused only slight or no reduction in heights of both regions. However, after the induction phase at this 0.4 mg/kg dosing, continued use of MTX at a low dose (once weekly at 0.2 mg/kg) caused a reduction in primary spongiosa height and bone volume on weeks 9 and 14, which was associated with an increased osteoclast formation and their bone surface density as well as a decreased osteoblast bone surface density in the primary spongiosa. Folinic acid supplementation was shown able to prevent the MTX effects in the primary spongiosa. Acute use of MTX can damage growth plate and primary bone at a high dose, but not at a low dose. However, long-term use of MTX at a low dose can reduce primary bone formation probably due to decreased osteoblastic function but increased osteoclastic formation and function, and supplementary treatment with folinic acid may be potentially useful in protecting bone growth during long-term low-dose MTX chemotherapy."

Folinic Acid(leucovorin) prevents damage to the bone marrow.  It may help by reducing the number of micronuclei.

"Low-dose MTX is able to inhibit other folate-dependent enzymes such as thymidylate synthase, which further blocks the de novo purine synthesis by directly inhibiting the activity of 5-aminooimidazole-4-carboxamide ribonucleotide transformylase, causing an increase in both intracellular and extracellular adenosine (an potent anti-inflammatory mediator), and an increase in cAMP"

No data was taken on Folinic Acid treatment alone.

A comparison of vitamin A and leucovorin for the prevention of methotrexate-induced micronuclei production in rat bone marrow.

"Methotrexate [is] a folate antagonist. In rats, methotrexate is known to induce micronuclei formation, leading to genetic damage, while vitamin A is known to protect against such methotrexate-induced genetic damage. Leucovorin (folinic acid) is generally administered with methotrexate to decrease methotrexate-induced toxicity.
We aimed to determine whether vitamin A and leucovorin differed in their capacity to prevent formation of methotrexate-induced micronuclei in rat bone marrow erythrocytes. The present study also aimed to evaluate the effect of combined treatment with vitamin A and leucovorin on the formation of methotrexate-induced micronuclei.
Male and female Wistar rats were injected with 20 mg/kg methotrexate (single i.p. dose). The control group received an equal volume of distilled water. The third and fourth groups of rats received vitamin A (5000 IU daily dose for 4 successive days) and leucovorin (0.5 mg/kg i.p. dose for 4 successive days), respectively. The fifth and sixth groups of rats received a combination of vitamin A and a single dose of methotrexate and a combination of leucovorin and methotrexate, respectively. The last group of rats received a combination of leucovorin, vitamin A and single dose of methotrexate. Samples were collected at 24 hours after the last dose of the treatment into 5% bovine albumin. Smears were obtained and stained with May-Grunwald and Giemsa. One thousand polychromatic erythrocytes were counted per animal for the presence of micronuclei and the percentage of polychromatic erythrocyte was determined.
Comparison of methotrexate-treated rats with the control group showed a significant increase in the percentage of cells with micronuclei and a significant decrease polychromatic erythrocyte percentage. Combined methotrexate and vitamin A therapy and combined methotrexate and leucovorin therapy led to significant decreases in the micronuclei percentage and an increase in polychromatic erythrocyte percentage when compared to rats treated with methotrexate alone. Leucovorin was found to be more effective than vitamin A against the formation of methotrexate-induced micronuclei."

"rats treated with a combination of methotrexate and vitamin A had a significantly reduced frequency of micronuclei formation when compared to methotrexate (20mg/kg)-treated rats"

Leucovorin helps to prevent genetic damage.  Does it do it in cases not caused by Methotrexate?

Inhibitory effect of folinic acid on radiation-induced micronuclei and chromosomal aberrations in V79 cells.

"Folinic acid (FA), clinically called leucovorin, has been widely used as a nutrient supplement in dietary intake and is capable of inhibiting cytotoxicity and chromosomal damage induced by chemicals. However, data on its antigenotoxic effect on radiation-induced chromosomal damage are limited. The present study was, therefore, performed to investigate the effect of FA on radiation-induced (X-rays and UV radiation) micronuclei (MN) and structural chromosomal aberrations (SCA) concurrently in V79 Chinese hamster lung cells. Exponentially growing cells were exposed to five doses of X-rays (1-12 Gy) and UV radiation (50-800 microJ x 10(2)/cm2) and post-treated with 5 or 50 micrograms FA/ml of culture medium for 16 h. The slides were analyzed for the presence of MN and SCA using standard procedures. X-ray treatment alone produced dose-related cytotoxicity. X-rays produced a clear dose-related clastogenicity as measured by percent of micronucleated binucleated cells (MNBN) (5-79%) and percent of aberrant cells (11-92%). FA at 5 micrograms/ml slightly decreased X-ray induced chromosomal damage in both assays; however, the inhibition was significant (12-46% of MNBN, 14-48% in aberrant cells) only when X-ray-treated cultures were post-treated with 50 micrograms FA/ml. Post-treatment of FA had no effect on X-ray induced cytotoxicity as measured by NDI and MI. A similar a dose-related increase in % MNBN (0.5-10.3%) and percent aberrant cells (6-35%) was produced by UV radiation treatment alone. There were significant percentages of MNBN and aberrant cell inhibitions at both 5 and 50 micrograms/ml in both assays. As in the case of X-ray-treated cells, there was a clear dose-related cytotoxicity in UV-treated cells alone. No reduction in NDI or MI was found when UV-exposed cells were post-treated with 5 or 50 micrograms of FA. FA [decreases] radiation-induced chromosomal damage."

"X-rays produce DNA double-strand breaks (DSBS), DNA single-strand breaks(SSBs), base damages and DNA-protein cross-links and that DSBs are the main lesions responsible for chromosome aberrations"

Folinic Acid protects against many sources of DNA damage(and DNA damage could include damage to DNA Methyltransferase explaining it's growth stunting properties).

Low expression of gamma-glutamyl hydrolase mRNA in primary colorectal cancer with the CpG island methylator phenotype.

"The CpG island methylator phenotype (CIMP+) in colorectal cancer (CRC) is defined as concomitant and frequent hypermethylation of CpG islands within gene promoter regions. We previously demonstrated that CIMP+ was associated with elevated concentrations of folate intermediates in tumour tissues[elevated folate levels may cause hypermethylation]. In the present study, we investigated whether CIMP+ was associated with a specific mRNA expression pattern for folate- and nucleotide-metabolising enzymes. An exploratory study was conducted on 114 CRC samples from Australia. mRNA levels for 17 genes involved in folate and nucleotide metabolism were measured by real-time RT-PCR. CIMP+ was determined by real-time methylation-specific PCR and compared to mRNA expression. Candidate genes showing association with CIMP+ were further investigated in a replication cohort of 150 CRC samples from Japan. In the exploratory study, low expression of gamma-glutamyl hydrolase (GGH) was strongly associated with CIMP+ and CIMP+-related clinicopathological and molecular features. Trends for inverse association between GGH expression and the concentration of folate intermediates were also observed. Analysis of the replication cohort confirmed that GGH expression was significantly lower in CIMP+ CRC. Promoter hypermethylation of GGH was observed in only 5.6% (1 out of 18) CIMP+ tumours and could not account for the low expression level of this gene. CIMP+ CRC is associated with low expression of GGH, suggesting involvement of the folate pathway in the development and/or progression of this phenotype. Further studies of folate metabolism in CIMP+ CRC may help to elucidate the aetiology of these tumours and to predict their response to anti-folates and 5-fluorouracil/leucovorin."

Folate is necessary for hypermethylation as Folate is a vitamin essential for DNA synthesis.  Although, excess levels of Folate will likely just be urinated out.

Folate and cancer: how DNA damage, repair and methylation impact on colon carcinogenesis.

"Inappropriate diet may contribute to one third of cancer deaths. Folates, a group of water-soluble B vitamins present in high concentrations in green, leafy vegetables, maintain DNA stability through their ability to donate one-carbon units for cellular metabolism. Folate deficiency has been implicated in the development of several cancers, including cancer of the colorectum, breast, ovary, pancreas, brain, lung and cervix. Generally, data from the majority of human studies suggest that people who habitually consume the highest level of folate, or with the highest blood folate concentrations, have a significantly reduced risk of developing colon polyps or cancer. However, an entirely protective role for folate against carcinogenesis has been questioned, and recent data indicate that an excessive intake of synthetic folic acid (from high-dose supplements or fortified foods) may increase human cancers by accelerating growth of precancerous lesions. Nonetheless, on balance, evidence from the majority of human studies indicates that dietary folate is genoprotective against colon cancer. Suboptimal folate status in humans is widespread. Folate maintains genomic stability by regulating DNA biosynthesis, repair and methylation. Folate deficiency induces and accelerates carcinogenesis by perturbing each of these processes."

Folic Acid (Nature Made Folic Acid Supplement, 400 mcg, 250-Count Tablets (Pack of 3)) as well as Folinic Acid can maximize height growth by protecting against DNA damage.  But excess folic or folinic acid are only necessary but not sufficient conditions for DNA Methylation to occur.  Every child undergoing growth should ensure that they have appropriate quantities of folic and folinic acid to maximize height growth.

DNA Hypermethylation needs something else to occur and one suggested mechanism is S-Adenosyl methionine.

Effect of methotrexate and folinic acid on skeletal growth in mice.

"Four equal groups of Balb/c young male mice (6 animals in each group; mean body weight 11.9 +/- 0.25 g, in their rapid growth phase[3 weeks]) were subjected to the following drug treatment for a period of 3 wk. Group 1 was given intraperitoneal MTX (3.5 mg kg(-1) body weight) every second day. Group 2 received folinic acid (7.0 mg kg(-1) body weight) intraperitoneally every second day. Group 3 was given both drugs (MTX every second day and folinic acid 8 h post-MTX injection). Group 4 was injected with physiological saline every other day to serve as a control group.

Mean lengths of both the tibia and femur of animals were compared in the four treatment groups. A significant decrease in the mean lengths was observed in the group receiving MTX alone. Similarly, there was a significant decrease in the height of the femoral and tibial growth plate in this group when compared with the other groups. The main effect of MTX seemed to be on the hypertrophic proliferative zone of chondrocytes in the growth plate. animals in this MTX-treated group also showed increased levels of MTX in plasma and low levels of erythrocyte folate{thus folate deficiency may play a role in the length decrease}.

Chronic administration of MTX induces suppression of skeletal growth in mice, possibly through the inhibition of the pathway of de novo DNA synthesis{thus other compounds that inhibit new DNA synthesis may also decrease height}. Folinic acid treatment following MTX administration appears to reverse this growth inhibition."

Folinic Acid alone non-significantly increased both femur and tibia length.

A is saline, B is folinic acid, C is MTX, D is MTX + folinic acid.  The growth plate quality seems higher in group B.

Same groups as above.  Again, GP with highest quality is B.  What's interesting to note is that in D the hypertrophic zone seems to be disorganized but the proliferative zone seems expanded.  Therefore, Folinic Acid may revert MTX growth inhibition by more an expansion of the proliferative zone rather than reverting all the effects of MTX.

Growth plate height was non-significantly higher in Folinic Acid group than in the other groups by almost 10% for the femur.  I think why it was shown as non-significant is that the p value chosen was very low p <0.001.

Folinic Acid increased folate levels in erythrocytes by about 40%.

Prevention of bone growth defects, increased bone resorption and marrow adiposity with folinic acid in rats receiving long-term methotrexate.

"chronic folinic acid supplementation can prevent methotrexate-induced chondrocyte apoptosis and preserve chondrocyte columnar arrangement and number in the growth plate. In the metaphysis, folinic acid supplementation can preserve primary spongiosa heights and secondary spongiosa trabecular volume by preventing osteoblasts from undergoing apoptosis and suppressing methotrexate-induced marrow adiposity and osteoclast formation. Systemically, plasma of folinic acid supplemented rats, in comparison to plasma from rats treated with MTX alone, contained a significantly lower level of IL-1β and suppressed osteoclast formation in vitro in normal bone marrow cells. The importance of IL-1β in supporting plasma-induced osteoclast formation was confirmed as the presence of an anti-IL-1β neutralizing antibody attenuated the ability of the plasma (from MTX-treated rats) in inducing osteoclast formation."

This study did not study Folinic Acid supplementation without MTX.

Comparative Histological Study on the Protective Effect of Folinic Acid versus Fish Oil on the Growing Bone of Methotrexate-Treated Young Rats

"Forty two, 6 weeks-old-male albino rats were divided into: group I (control), group II (MTXtreated), group III (MTX and FA-treated) and group IV (MTX and FO-treated). MTX was injected subcutaneously, once daily for 5 consecutive days, 0.65 mg/kg, followed by 9 days of rest, then 1.3 mg/kg twice weekly for 4 weeks. FA was injected intraperitoneally, 6 hours after each dose of MTX, 0.87 mg/kg, then1.3 mg/kg twice weekly. FO was given orally daily for 6 weeks, 0.5 ml/100 gm. Left knee joints were processed for measuring RANKL/ OPG ratio (Receptor Activator of Nuclear factor Kapp-B Ligand/ Osteoprotegerin). Right knee joint sections were stained with H&E, Masson's Trichrome and immunohistochemical staining for Caspase-3. Morphometric measurements and statistical analysis were done. Results: MTX-treated group sections revealed disruption in the growth plate structure with subsequent reduction in endochondral bone formation. Supplementation with FA and FO preserved growth plate integrity and bone formation."

"Mean values of rats' length in cm (±SD) in the studied groups Group Mean ± SD Group I (control) 23.34±0.5 Group II 14.34±0.37*□ Group III 21.26±0.79*• Group IV 22.67±0.45•"<unfortunately there was no folinic acid only group

LSJL increases DNA Methylation

Since DNA Methylation markers tend to be upregulated during cellular differentiation, this is further evidence that LSJL induces chondrogenic differentiation.

Endothelin is upregulated 2.258 fold by LSJL.

Localized methylation in the key regulator gene endothelin-1 is associated with cell type-specific transcriptional silencing.

"To identify key developmental regulator genes whose expression in terminally differentiated cells may be inhibited by DNA methylation, mouse dermal fibroblasts were demethylated with 5-aza-2'-deoxycytidine, and changes in gene expression monitored. Endothelin-1 (Et1 or Edn1), which encodes a cytokine with diverse regulatory functions, was among the genes upregulated following demethylation. CpG dinucleotides within a short region in intron 1 of the gene have dramatically higher levels of methylation in Et1-non-expressing fibroblasts and chondrocytes as compared to the Et1-expressing mouse cell line, mIMCD-3. Strong evolutionary conservation of this region implies its role in the cis-regulation of Et1 transcription. To confirm that should Et1 in dermal fibroblasts become aberrantly activated, it could indeed lead to the dysregulation of many downstream genes, we exposed fibroblasts to exogenous ET1 peptide and assayed for transcriptional changes by microarray. ET1 treatment resulted in significant expression changes - primarily downregulation - of a significant number of genes. In particular, Tgfbeta2 and Tgfbeta3 were among the downregulated genes, which in turn alter the expression status of their many target genes."

"inhibition of Et1 in fibroblasts and chondrocytes may involve targeted methylation of the Sp1 binding site (among other sites) in intron 1, which block Sp1 binding and contribute to Et1 silencing."

New born mice were used.

Bold genes were also altered in demethylated fibroblasts.

Genes upregulated in Fibroblasts that were treated with exogenous et1 that also upregulated(or downregulated) in LSJL:

MT1

MT2

Car6

Slc7a3(downregulated in LSJL)

Genes downregulated in exogenous et1 fibroblasts:

Asporin(up in LSJL)

cnn1(up in LSJL)

col6a2(up in one DNA region down in another)

Diap3

Prrx2(up in LSJL)

cxcl5

ccnb2

col6a1(up in LSJL)

c1qtnf3(up in LSJL)

slc38a4(up in LSJL)

Ccl2(up in LSJL)

ccl7(up in LSJL)

il6(up in LSJL)

9930013L23Rik(up in LSJL)

Lmo7
Lrrc15(up in LSJL)

A large portion of differentially regulated genes were genes that were altered in demethylated fibroblasts.  A large portion of the similarly regulated genes were those only effected by Et1 which corresponds to the data as knee loading upregulated Et1.  So LSJL likely involves cellular methylation.


Epigenomic and microRNA-mediated regulation in cartilage development, homeostasis, and osteoarthritis.

"epigenetic derepression [IS] associated with DNA methylation loss on chondrocyte genes, including MMP3[up in LSJL], MMP9, MMP13, ADAMTS4[up in LSJL], IL-1β, and LEP"

5-Azacytidine makes human preadipocytes able to differentiate into mesoderm-derived cell lineages.

"In the present study we have evaluated whether (i) 5-azacytidine (AZA), a well-known demethylating agent, could be able to modify the phenotype of human preadipocytes and (ii) the modified cells could possess multilineage differentiation potential. Human preadipocytes at the 3rd passage were treated for 48 or 96 h with 10 μM AZA and then expanded up to passage 5. Stem cell markers, such as OCT-4, Nanog, and Sox2, were upregulated after 96 h of treatment with the demethylating treatment. Further, decreases in the expression of genes, such as adipose differentiation-related protein, characterizing the preadipocytes were noted. AZA-treated preadipocytes differentiated into cell lineages derived from mesoderm. Indeed, after incubation with inductive media for 3 weeks, osteblast-, chondrocyte-, and myoblast-like cells were detected in the cultures. Interestingly, both upregulation of stem cell markers and differentiation potential were maintained by the treated cultures expanded until the 5th passage. AZA, without the use of transduction methods, convert preadipocytes to a less differentiated state that can be induced, under suitable stimuli, to the formation of mesoderm-derived cell lineages."

Instead of trying to induce methylation you may want to induce demethylation to induce chondrogenesis.  Since LSJL increases methylation, decreasing methylation in the bone before LSJL and only then(as decreasing methylation will reduce differentiation as well) may allow for more available stem cells to differentiate chondrocytes.  However, the next study suggests that the methylation status of chondrogenic genes is already low in adult human cells.

Methylation status of CpG islands in the promoter regions of signature genes during chondrogenesis of human synovium-derived mesenchymal stem cells.

"DNA methylation is one mechanism that regulates human chondrogenesis.  [What's] the CpG methylation status in human synovium-derived MSCs during experimental chondrogenesis?

Human synovium-derived MSCs were subjected to chondrogenic pellet culture for 3 weeks. The methylation status of 12 regions in the promoters of 10 candidate genes (SOX9, RUNX2, CHM1, FGFR3, CHAD, MATN4, SOX4, GREM1, GPR39, and SDF1) was analyzed before and after differentiation. The expression levels of these genes were analyzed. Methylation status was also examined in human articular cartilage.

10 of the 11 CpG-rich regions analyzed were hypomethylated in human progenitor cells before and after 3 weeks of pellet culture, regardless of the expression levels of the genes. The methylation status was consistently low in SOX9{up in LSJL}, RUNX2, CHM1, CHAD, and FGFR3 following an increase in expression upon differentiation and was low in GREM1 and GPR39 following a decrease in expression upon chondrogenesis. One exceptional instance of a differentially methylated CpG-rich region was in a 1-kb upstream sequence of SDF1, the expression of which decreased upon differentiation. Paradoxically, the hypermethylation status of this region was reduced after 3 weeks of pellet culture.

The DNA methylation levels of CpG-rich promoters of genes related to chondrocyte phenotypes are largely kept low during chondrogenesis in human synovium-derived MSCs."

"chondromodulin 1 (CHM1), fibroblast growth factor receptor 3 (FGFR3), [MATN4]{up in LSJL}, and chondroadherin (CHAD) for genes up-regulated in chondrogenic pellet cultures, and for genes down-regulated in chondrogenic pellet cultures, these were SOX4, Gremlin 1 (GREM1), G protein–coupled receptor 39 (GPR39), and stromal cell–derived factor 1 (SDF1)."

"DNA methylation conditions in immature mesenchymal cells are permissive of the expression of chondrocyte phenotype"

Although one study suggests that loss of DNA methylation is associated with cellular senescence.  However this may have been only a correlational association and not a causal association.  Also, DNA methylation may be a marker of cellular differentiation state which corresponds to other data rather than a cell cycle differentiation counter.

It's possible that the upregulation of DNA Methylation related genes is due to fibroblast differentiation.  However the presence of chondrogenic genes like Sox9 confirms condrogenic differentiation. 

Increase your Height with S-Adenosyl methionine

Earlier, we discussed how DNA Methylation was involved in growth plate cessation.  Yes, Estrogen handles growth plate fusion but growth plate fusion will not occur until after growth plate cessation.  Then we discussed how folic and folinic acid were involved in producted DNA from damage(And in turn damage to DNA Methyltransferase).  S-Adenosyl Methionine is available for purchase:Nature Made SAM-e Complete 200 mg - 90 Enteric Coated Tablets

Now, S-Adenosyl Methionine has a methyl group that can be donated to different proteins.  If methyl groups were continuously donated to growth plate chondrocytes the body would keep growing!

Exploring the mechanisms behind S-adenosylmethionine (SAMe) in the treatment of osteoarthritis

"Clinical trials have shown reduced pain and stiffness [in osteoarthritis] while in vitro and animal studies have shown SAMe can stimulate the production of cartilage which is critical in reversing the disease process. The author examines many potential mechanisms of action including: reduction of inflammatory mediators; increasing levels of glutathione; direct or indirect signaling of cartilage synthesis or survival; maintenance of DNA methylation. Research into the mechanisms of supplemental SAMe in osteoarthritis is necessary to evaluate the clinical effectiveness and safety of this dietary supplement."

"Osteoarthritic chondrocytes are particularly sensitive to the inflammatory cytokines because they have increased levels of their activating enzymes and receptors. Part of the catabolic role of IL-1 could be due to its ability to up regulate the inducible form of nitric oxide synthase and cyclooxygenases (COX)-2, leading to the increased production of nitric oxide and prostaglandin E2. Nitric oxide can induce apoptosis and decrease proteoglycan synthesis in chondrocytes by interfering with β 1-integrin mediated cell to matrix signal transduction mechanism. Over time, the cartilage would disintegrate, resulting in narrowing of the joint space and damage to the surrounding bone"

"joint space narrowing occurs at < 0.1 mm per year"

"intramuscular injection with 30 mg/kg and 60 mg/kg of SAMe for 12 weeks resulted in an increased number of cells and depth of cartilage in a partial meniscectomy model of osteoarthritis in rabbits compared to placebo"

"positive effects after treatment with both 1 μ g/ml and 10 μ g/ml SAMe, with 10 μ g/ml being the most effective at increasing proteoglycan synthesis and secretion, while 100 μ g/ml had an inhibitory effect"

"SAMe plays a vital role in three biochemical pathways; methylation, transsulfuration and aminopropylation. The loss of a methyl group from SAMe triggers the transsulfuration pathway creating all endogenous sulfur compounds. With the loss of its methyl group to different acceptors (i.e. DNA, proteins, phospholipids, and neurotransmitters) SAMe is converted into s-adenosylhomocysteine, which is hydrolyzed to adenosine and homocysteine. When methionine is needed, homocysteine is remethylated by methyltetrahydrofolate homocysteine methyltransferase with B12 as a cofactor. When methionine is in excess, homocysteine is converted to cystathionine by β cystathionine-synthase with vitamin B6 as a cofactor. Cystathionine is then converted to cysteine, which can act as a reducing agent either alone or as an active part of glutathione. Polyamines are synthesized via the aminopropylation pathway which begins when SAMe is converted to decarboxylated SAMe via SAMe decarboxylase. As an aminopropyl group is transferred to putrescine, methylthioadenosine is formed, which is followed by the production of spermidine and spermine. Methylthioadenosine is then converted back to methionine"

Well, in osteoarthiritis the cartilage is already turning into bone so we don't know if SAMe consumption will result in endochondral ossification of the articular cartilage.  But, SAMe can stimulate the production of cartilage when ordinarily cartilage production is very limited so taking SAMe during development could lead to extra height growth(If you take SAMe you also need to take Folic acid as well to cope with the additional cellular proliferation).  SAMe is produced by the body.

Moderate or supranormal folic acid supplementation does not exert a protective effect for homocysteinemia and methylation markers in growing rats

"Folic acid (FA) deficiency/supplementation effects seem to be dependent on age group and/or physiological status. The aim was to evaluate changes associated with rapid growth in relation to methionine metabolism in rats.
Four groups (n = 10 each) of male Sprague Dawley rats (5 weeks old) were on diets that varied in their FA content: 0 mg FA/kg diet (deficient), 2 mg FA/kg diet (control), 8 mg FA/kg diet (moderate supplementation), 40 mg FA/kg diet (supranormal supplementation). Animals were fed ad libitum for 30 days. Biomarkers of methionine metabolism and antioxidant status were evaluated.
Serum total homocysteine concentration increased (p < 0.01) in FA deficient animals, with no differences between the supplemented groups. The hepatic 'methylation ratio' (S-adenosylmethionine/S-adenosylhomocysteine) of the FA content groups reached similar values, which were significantly higher compared to the deficient group. The brain 'methylation ratio', however, remained unmodified independently of FA content in the diet. FA deficiency induced hepatic DNA hypomethylation, and supranormal FA supplementation exerted the most protective effect (p < 0.01). Serum folate levels increased according to FA dietary level, whereas no differences were seen for vitamin B(12) and vitamin B(6).
FA deficiency compromises methionine metabolism whereas supplementation does not show an additional positive effect compared to the control diet in growing animals"

"The main dietary components that act as methyl groups donors are folates, methionine, vitamin B 12 and choline"

"The methionine cycle is nutritionally regulated by FA, and vitamins B 6 and B 12"

"The so-called methylation ratio (AdoMet/AdoHcy) of the 3 groups whose diet included FA reached similar values, significantly higher compared to the deficient group"

"Serum folate levels are increased, as expected, in accordance with the vitamin supplementation level of the diet. However, serum vitamin B 6 and B 12 in the different groups remained unmodified regardless of dietary FA content"

"neither FA deficiency nor supplementation altered the average weight gain of the animals
throughout the study. These results are comparable to other studies that show neither deficiency nor supplementation of FA in the diet inhibits or improves animal response in relation to growth"

After 200 micrograms/a day of FA, Folic acid is no longer metabolized and floats in the serum.

"high-dose FA supplementation does not seem to induce higher transmethylation activity"

The conclusions seem to differ from the results as they say that supranormal FA supplementation exerts a protective effect and serum folate levels increased with Folic acid supplementation.  But this shows that SAMe supplementation will have an effect independent of Folic acid supplementation as in all the groups the SAMe level in the blood normalized on a particular level.

SAMe hasn't really been tested on healthy adults and it's effects on cell growth but in theory SAMe supplementation should result in some extra height growth as a result of more chondrocytes getting methylating and thus proliferating and differentiating for a longer period.

Increase Height with DNA Methylation?

Previously, in the blog I reported the function of osteoclasts in growing taller.  The use of alendronate inhibited osteoclasts reobsorption capacity and that reduced overall height growth.  What's interesting is that the growth plate was actually larger when osteoclasts were inhibitied.  The authors suggested that this was a result of failure of VEGF to induce chondrocyte apoptosis.  It was also reported previously that growth plate senescence is a result of a decrease in DNA Methylation(Senescence being when a growth plate is inactive).

DNA Methylation is the addition of a Methyl Group to DNA and affects DNA regulation(so transcriptional proteins that stop growth can no longer bind to the gene). DNA Methylation also play a role in cell memory(so cells that were once chondrocytes no longer remember that they're supposed to be chondrocytes).

How does DNA Methylation affect height growth and how does it relate to osteoclasts? Are there any chemicals or substances we can use to alter DNA Methylation within the body?

Expression profile of genes related to osteoclastogenesis in mouse growth plate and articular cartilage.

"Cartilage tissue is broadly classified into transient cartilage (e.g. growth plate, GP) and permanent cartilage (e.g. articular cartilage, AC). The former eventually disappears and is replaced by bone during the endochondral ossification process, whereas the latter retains its permanency. Osteo(chondro)clasts, multinucleated giant cells of the monocyte/macrophage lineage, are selectively induced in the GP during endochondral ossification and play central roles in the resorption of cartilagenous matrices. The aim of this study was to investigate the factors determining the GP-specific recruitment of osteo(chondro)clasts. We especially focused on the expression pattern of the receptor activator of NF-kappaB ligand (RANKL), an essential factor for osteo(chondro)clast differentiation, and on that of epigenetic and transcriptional factors affecting RANKL gene expression. Knee joints of male BALB/c mice aged 8 weeks were dissected and subjected to immunohistochemical analysis using anti-RANKL, Runx2, Dlx5 and Msx2 antibodies. The methylation status of the mouse RANKL gene promoter in both the GP and the AC was analyzed. The expression of BMP-2, -3, -4, -6 and type X collagen mRNA was examined. At the boundary between the calcifying cartilage and the hypertrophic chondrocytes of the GP, RANKL-expressing chondrocytes overlapped those expressing Runx2, Dlx5 and Msx2, near numerous osteo(chondro)clasts. Although similar BMP-2 and -4 expression was observed in chondrocytes in both the GP and the AC as well as in maturing osteoblasts, a rather restricted BMP-6 expression pattern was observed in resting and proliferating chondrocytes in the GP. Mostly non-CpG methylation was scattered in a non-specific manner in chondrocytes in the GP and the AC. Putative Runx2 binding elements are located in the RANKL promoter. Runx2, an essential transcription factor for skeletal development, is a key regulator of RANKL expression in chondrocytes in the GP."

DNA Methylation in growth plates is linked to osteoclasts by RANKL which activates osteoclasts.  BMP-6 is a difference between that chondrocytes of the Articular Cartilage and Growth Plate but if endochondral ossification can be performed safely and effectively within the articular cartilage is unknown.  Osteoclasts may absorb catiligenous matrices which prevent cartilage cells from reaching their full potential.

Growth plate senescence is associated with loss of DNA methylation

"One mechanism that has been proposed to explain replicative senescence involves epigenetic changes in methylation of genomic DNA. Some CG sequences in mammalian genomic DNA are methylated on the cytosine moiety. When DNA is replicated, the new strand is initially not methylated. DNA methyltransferase 1, a maintenance methylase, recognizes the hemimethylated CGs and adds the missing methyl groups. If maintenance methylation is incomplete, methylation levels may gradually decrease with repeated cell replication.  The level of DNA methylation could serve as a cell-cycle counter. The level of DNA methylation can then affect gene expression, in part by altering interaction of transacting elements with the promoter region and in part by altering histone modification, and thus chromatin structure. Such epigenetic changes may contribute to replicative senescence and terminal differentiation in some cell types. Growth plate chondrocytes cultured at high density and exposed to a demethylating agent (5-azacytidine), undergo hypertrophic differentiation. Disruption of PASG (proliferation-associated SNF2-like gene), PASG, which is required for normal maintainance of DNA methylation, results in growth retardation and premature aging. "

So if DNA Methylation does not occur then there is growth retardation but DNA Methylation may serve as a cell-cycle counter to tell growth plates when to stop. So what we have to do is set back the cell-cycle counter some steps to trick it into thinking the number of divisions it's done is less than it currently is.

"Growth plate chondrocytes cultured at high density and briefly exposed to 5-azacytidine, a demethylating agent, differentiate and start expressing markers specific for hypertrophic chondrocytes"<-Since methylation markers increase with differentiation, demethylating chondrocytes may trick them into thinking that they're at an earlier differentiation state.

"We first tested the prediction that cultured growth plate chondrocytes from older animals will undergo fewer cell divisions before undergoing replicative senescence than will chondrocytes from younger animals. Resting zone chondrocytes extracted from fetal, 4-, and 16-week-old male rabbits proliferated for approximately 50 days and underwent three passages (with increasing intervals) before reaching senescence. During this time, chondrocytes from animals of different ages underwent a similar number of population doublings (13.1 ± 1.1 vs 14.6 ± 0.6 vs 14.3 ± 0.8; fetal vs 4 weeks vs 16 weeks respectively). Chondrocytes in primary culture were small, had a round or polygonal shape, and stained for alcian blue and alkaline phosphatase activity, but not for senescence related ß-galactosidase. In contrast, chondrocytes that were becoming senescent were larger, only stained faintly or not at all for alcian blue and alkaline phosphatase activity, but showed an increased senescence-related ß-galactosidase activity"<-Senesent chondrocytes had low alkaline phosphatase level so that may be key for growth. A Senesence B-Galactosidase inhibitor might also increase growth.

"[The] loss of DNA methylation could occur as these resting zone chondrocytes gradually replicate if there is incomplete maintenance methylation of the newly synthesized strand of DNA.  Growth plate senescence is associated with a loss of DNA methylation in rib resting zone chondrocytes. Loss of DNA methylation was observed in the resting zone chondrocytes of the distal ulna. In the distal ulna, loss of methylation with age was observed in the proliferative and hypertrophic zone chondrocytes which are thought to be progeny of the resting zone chondrocytes. Within each age, there was no significant difference in the level of DNA methylation between the different zones of the growth plate. Loss of methylation appears to occur specifically during replication of resting zone chondrocytes but not during the more rapid proliferation of proliferative zone chondrocytes.  There may be complete maintenance methylation in the proliferative zone, but not in the resting zone. Loss of methylation might be responsible for the temporal limits that cause chondrocyte replication to slow with age but not the spatial limits that cause chondrocyte proliferation to slow as the cells descend farther down the chondrocyte columns. The spatial limitation on proliferation may not be controlled by a cell-cycle counter, but instead it may be controlled by a chemical gradient, e.g. parathyroid hormone-related protein."

So you have to make sure those resting zone chondrocytes(stem cells) stay methylated.  I don't know of any good DNA Methylation stimulators.

According to SOX trio decrease in the articular cartilage with the advancement of osteoarthritis, there was no association between age and a reduction of SOX9 methylation promoter regions.

Epigenetic modifiers influence lineage commitment of human bone marrow stromal cells: Differential effects of 5-aza-deoxycytidine and trichostatin A.

"We investigated the effects of the DNA demethylating agent 5-aza-2'-deoxycytidine (5-aza-dC) or the histone deacetylase inhibitor trichostatin A (TSA) on osteogenic and chondrogenic differentiation. Monolayer cultures of HBMSCs were treated for 3 days with the 5-aza-dC or TSA, followed by culture in the absence of modifiers. Cells were subsequently grown in pellet culture to determine matrix production. 5-aza-dC stimulated osteogenic differentiation as evidenced by enhanced alkaline phosphatase activity, increased Runx-2 expression in monolayer, and increased osteoid formation in 3D cell pellets. In pellets cultured in chondrogenic media, TSA enhanced cartilage matrix formation and chondrogenic structure. epigenetic modifiers, as agents, possibly in combination with other factors,  enhance the ability of HBMSCs to form functional bone or cartilage with significant therapeutic implications therein."

In the study TSA increased COL2A1 and Aggrecan levels which are Sox related genes.

So the supplement that encourage demethylation enhanced osteogenic differentiation.  So this indicates that perhaps supplements that encourage DNA methylation may have the reverse effect(chondrogenic differentiation).

Epigenetic regulation in chondrogenesis.

"DNA methylation in CpG-rich promoters correlates with gene silencing. Histone modification including histone acetylation and deacetylation determines the stability of the chromatin structure. Condensed chromatin (heterochromatin), which has a higher-order histone-DNA structure, prevents the access of transcriptional activators to their target genes. The fundamental unit of eukaryotic chromatin consists of 146 bp of DNA wrapped around a histone octamer. Posttranslational modifications of the histone tail and the chromatin remodeling complex disrupt histone-DNA contacts and induce nucleosome mobilization. Histone acetylation of specific lysine residues in the histone tail plays a crucial role in epigenetic regulation. Histone acetylation is a dynamic process regulated by the antagonistic actions of 2 families of enzymes - the histone acetyltransferases (HATs) and the histone deacetylases (HDACs). The balance between histone acetylation and deacetylation serves as a key epigenetic mechanism for transcription factor-dependent gene expression and the developmental process. DNA methylation, histone acetylation modified by HAT and/or HDAC, and transcription factor-associated molecules contribute to a mechanism that can alter chromatin structure, gene expression, and cellular differentiation during chondrogenesis."

"In chondrocyte differentiation, TGF-β stimulation is necessary for MSC-derived primary chondrogenesis. On the other hand, chondrocyte maturation is inhibited by TGF-β. These conflicting effects of TGF-β during chondrogenesis might depend on chromatin structure and/or the epigenetics of each differentiated stage."

"TGF-β-regulated Smad3 activates the Sox9-dependent transcription on the chromatin structure"

"CpG-rich promoters of chondrogenic-related genes, such as Sox9, Runx2, chondromodulin-I, and fibroblast growth factor receptor 3, are hypomethylated during synovium-derived chondrogenesis"<-Hypomethylated means undermethylated so perhaps too much methylation is bad for chondrogenesis.

"The Col2a1 gene is less methylated in chondrocytes than in fibroblasts"

"The demethylation of the Col10a1 promoter correlates with Col10a1 induction during MSC-derived chondrogenesis"

"In chondrogenesis, p300 stimulates transcription factor-mediated chromatin disruption. Coactivator p300 directly associates with the master chondrogenic factor Sox9, and activates Sox9-dependent transcription. Sox9-dependent transactivation is induced by p300-mediated histone acetylation of chromatin"

"p300 potentiates Sox9-dependent transcription on a chromatinized DNA template and is associated with hyperacetylated histones"

"Coactivator Tip60, which mainly acetylates H4, increases Sox9/Sox5-dependent Col2a1 transcription by associating with Sox9 on chromatin"

"HDAC4, which is expressed in prehypertrophic chondrocytes, regulates chondrocyte hypertrophy and endochondral bone formation by interacting with and inhibiting the activity of Runx2"

"Growth factors, cytokines, and nonproteinaceous chemical compounds including dexamethasone, vitamin D3, prostaglandin E2, and ascorbic acid influence gene expression and cellular differentiation during chondrogenesis"

"BMP-2 induces histone hyperacetylation and methylation at the Sox9 gene on chromatin"

So perhaps you actually want less methylation in genes related to chondrogenesis so a supplement like Sam-e wouldn't help increase height.


Genetic and non-genetic influences during pregnancy on infant global and site specific DNA methylation: role for folate gene variants and vitamin B12.

"[Folate metabolism has a] central role in provision of methyl groups for DNA methylation. Global (LUMA) and gene specific (IGF2, ZNT5, IGFBP3) DNA methylation were quantified in 430 infants. Seven polymorphisms in 6 genes (MTHFR, MTRR, FOLH1, CβS, RFC1, SHMT) involved in folate absorption and metabolism were analysed in DNA from both infants and mothers. Red blood cell folate and serum vitamin B(12) concentrations were measured as indices of vitamin status. Relationships between DNA methylation patterns and several covariates viz. sex, gestation length, maternal and infant red cell folate, maternal and infant serum vitamin B(12), maternal age, smoking and genotype were tested. Length of gestation correlated positively with IGF2 methylation[length of gestation means time it took for growth to occur, IGF2 methylation made growth take longer, larger animals have a larger gestation period] and inversely with ZNT5 methylation.  Methylation of the IGFBP3 locus correlated inversely with infant vitamin B(12) concentration[IGFBP3 decreases height], whilst global DNA methylation correlated inversely with maternal vitamin B(12) concentrations. Analysis of common genetic variants in folate pathway genes highlighted several associations including infant MTRR 66G>A genotype with DNA methylation and maternal MTHFR 677C>T genotype with IGF2 methylation. Both environmental and genetic factors involved in one-carbon metabolism influence DNA methylation in infants. Vitamin B(12) status, infant MTRR genotype and maternal MTHFR genotype, may influence the supply of methyl groups for DNA methylation. Gestational length [may determine] infant DNA methylation patterns."

"Reduced methylation at the IGF2 differentially methylated region, H19 DMR, in cord blood DNA has been associated with increased folic acid intake during pregnancy"<-IGF2 methylation could increase height and therefore folic acid intake during pregnancy may be contraindicated for height growth.

"maternal peripheral blood DNA methylation at the IGF2 locus was associated with maternal serum vitamin B12 levels"<-instead you'd want to take vitamin B12.


Bone morphogenetic protein-2 induces chromatin remodeling and modification at the proximal promoter of Sox9 gene.

"BMP-2 [induces] alterations in chromatin organization around the Sox9 core promoter. Nuclease hypersensitive site mapping following BMP-2 stimulation showed an inducible hypersensitive site in the Sox9 proximal promoter. BMP-2 increased the association of the transcription factor NF-Y with histone acetyltransferase p300/CBP.  The binding of the NF-Y-p300 complex to the Sox9 gene proximal promoter along with PCAF and RNA polymerase II. BMP-2 stimulation caused histone hyperacetylation and methylation at the Sox9 gene{But is Sox9 methylation a way to induce height growth or is it a marker that no further stimulation should be applied to Sox9?}. The activation of Sox9 gene transcription by BMP-2 is associated with chromatin remodeling and histone modification."

"BMP-2 regulates the chromatin structure of the Sox9 promoter through the p38 pathway, independent of the Smad pathway"


Epigenetic regulation of mesenchymal stem cells: a focus on osteogenic and adipogenic differentiation.

"The H3K27me3 mark is thought to be critical to the “stemness” of stem cells, as H3K27 demethylation triggers cellular differentiation"

"the ability of HMTs to methylate H3K9 in order to silence transcription often depends on the methylation status of adjacent lysine residues on H3"

"Acetylation of H3K9 (H3K9ac) and acetylation of H4K16 (H4K16ac) are common marks found on euchromatin near genes that are actively being transcribed"

"histone modification-mediated epigenetic alterations in late-passage MSCs may be responsible for a deceased ability to differentiate as cultured MSCs age."

"strong methylation of lineage specification and developmental promoters may restrict MSC differentiation capacity"


Epigenetic Regulation during Fetal Femur Development: DNA Methylation Matters.

" Using human embryonic stem cells, human fetal bone cells (HFBCs), adult chondrocytes and STRO-1(+) marrow stromal cells from human bone marrow, we have examined a spectrum of developmental stages of femur development and the role of DNA methylation therein. Using pyrosequencing methodology we analysed the status of methylation of genes implicated in bone biology; furthermore, we correlated these methylation levels with gene expression levels. During fetal femur development DNA methylation inversely correlates with expression of genes including iNOS (NOS2) and COL9A1{up}, but not catabolic genes including MMP13 and IL1B. Furthermore, significant demethylation was evident in the osteocalcin promoter between the fetal and adult developmental stages. Increased TET1 expression and decreased expression of DNA (cytosine-5-)-methyltransferase 1 (DNMT1) in adult chondrocytes compared to HFBCs could contribute to the loss of methylation observed during fetal development."

"COL9A1 expression levels were significantly correlated with fetal foot length"  Degree of Col9a1 methylation inversely correlated with foot length.

" the degree of [iNos] methylation was inversely correlated to foot length"

"[MMP13] expression significantly correlated with femur length"  Methylation of MMP13 is slightly inversely correlation with foot length.

Foot length is correlated with IL1B Methylation.

DNMT1 expression levels decrease with chondrocyte age.  There is much less of a correlation with age of TET1.