Comparison of Fibrocartilage to LSJL gene expression

Fibrocartilage is a mixture of fibrous tissue and cartilage.  Formation of fibrocartilage could be a sign of preliminary ectopic growth plate formation via LSJL.

Discrimination of meniscal cell phenotypes using gene expression profiles.

"The purpose of this study was to identify quantifiable characteristics of meniscal cells and thereby find phenotypical markers that could effectively categorize cells based on their tissue of origin (cartilage, inner, middle, and outer meniscus). The combination of gene expression ratios collagen VI/collagen II, ADAMTS-5/collagen II, and collagen I/collagen II was the most effective indicator of variation among different tissue regions."

The meniscus is a fibrocartilaginous structure.

"Although the meniscus is primarily composed of collagen I, the inner region shows a higher content of collagen II, the major component and indicator of hyaline cartilage"<-Both collagen types are upregulated by LSJL.

Genes upregulated in the meniscus versus articular cartilage also upregulated in LSJL:

Col1a1

Col6a1

Downregulated:

Col2a1{up}

Aggrecan{up}

Neutral(no obvious trends):

Versican{up}

Bgn{up}

ADAMTS4{up}

Cartilage intermediate layer protein 2 (CILP-2) is expressed in articular and meniscal cartilage and down-regulated in experimental osteoarthritis.

"Cilp2 is expressed differentially by articular chondrocytes. CILP-2 is highly homologous to CILP-1 (cartilage intermediate layer protein 1), which is expressed in the intermediate zone of articular cartilage. Cilp2 has a restricted mRNA distribution at the surface of the mouse articular cartilage during development, becoming localized to the intermediate zone of articular cartilage and meniscal cartilage with maturity. Although the extracellular CILP-2 protein localization is broadly similar to CILP-1, CILP-2 appears to be more localized in the deeper intermediate zone of the articular cartilage extracellular matrix at maturity. CILP-2 was shown to be proteolytically processed, N-glycosylated, and present in human articular cartilage. In surgically induced osteoarthritis in mice, Cilp1 and Cilp2 gene expression was dysregulated. However, whereas Cilp1 expression was increased, Cilp2 gene expression was down-regulated demonstrating a differential response to mechanically induced joint destabilization. CILP-2 protein was reduced in the mouse osteoarthritic cartilage. Ultrastructural analysis also suggested that CILP-2 may be associated with collagen VI microfibrils and thus may mediate interactions between matrix components in the territorial and inter-territorial articular cartilage matrix."

"The N-terminal of CILP-1 has been shown to bind to and inhibit TGFβ1 in vitro, and CILP1 mRNA is induced by TGFβ1. CILP-1 levels increase with age."

Cilp2 is not expressed in growth plate cartilage.

"The absence of Cilp2 mRNA and protein in growth plate cartilage suggests that CILP-2 is a component of permanent cartilage, rather than transient cartilage that will undergo ossification, and further defining the specialized composition of the extracellular matrix synthesized by articular chondrocytes."

Dynamic tensile loading improves the functional properties of mesenchymal stem cell-laden nanofiber-based fibrocartilage.

"we employ scaffolds composed of co-aligned nanofibers that direct mesenchymal stem cell (MSC) orientation and the formation of organized extracellular matrix (ECM). Concomitant with ECM synthesis, the mechanical properties of constructs increase with free-swelling culture, but ultimately failed to achieve equivalence with meniscal fibrocartilage.  This work examined the effect of cyclic tensile loading on MSC-laden nanofibrous constructs. Fiber-aligned scaffolds were seeded with MSCs and dynamically loaded daily in tension or maintained as nonloaded controls for 4 weeks. With mechanical stimulation, fibrous gene expression increased, collagen deposition increased, and the tensile modulus increased by 16% relative to controls."

"Four weeks of loading resulted in a two-fold increase in collagen I expression compared to nonloaded controls, with no effect on collagen II expression"

"Fibronectin, which is responsible for cell binding to the surrounding ECM and is a precursor to collagen deposition, increased by two-fold with conditioning. Likewise, lysyl oxidase{up in LSJL}, an enzyme that cross-links collagen fibrils, was significantly upregulated in loaded constructs compared to nonloaded controls"

Fibrocartilage tissue engineering: the role of the stress environment on cell morphology and matrix expression.

"Fibrocartilage develops at tendon-to-bone attachments and in compressive regions of tendons. Here, we developed and applied an in vitro system to determine whether fibrocartilage can develop under a state of periodic hydrostatic tension in which only a single principal component of stress is compressive. Mesenchymal stromal cells in a 3D culture were exposed to compressive and tensile stresses as a result of an external tensile hydrostatic stress field. Tensile cyclic stresses promoted spindle-shaped cells, upregulation of scleraxis{up} and type one collagen, and cell alignment with the direction of tension. Cells experiencing a single compressive stress component exhibited rounded cell morphology and random cell orientation. No difference in mRNA expression of the genes Sox9 and aggrecan was observed when comparing tensile and compressive regions unless the medium was supplemented with the chondrogenic factor transforming growth factor beta3. In that case, Sox9 was upregulated under static loading conditions and aggrecan was upregulated under cyclic loading conditions. Yhe fibrous component of fibrocartilage could be generated using only mechanical cues, but generation of the cartilaginous component of fibrocartilage required biologic factors in addition to mechanical cues."

In vitro study of stem cell communication via gap junctions for fibrocartilage regeneration at entheses.

"Entheses are fibrocartilaginous organs that bridge ligament with bone at their interface and add significant insertional strength. T

Rabbit bone/ligament fibroblasts were dual-stained with DiI-Red and calcein (gap-junction permeable dye), and cocultured with unlabeled BMSCs at fixed ratio (1:10). The functional gap junction was demonstrated by the transfer of calcein from donor to recipient cells that was confirmed and quantified by flow cytometry. Type 2 collagen (cartilage extracellular matrix-specific protein) expressed by the mixed cell lines in the cocultures were estimated by real-time reverse transcription PCR and compared with that of the ligament-bone coculture (control).

Significant transfer of calcein into BMSCs was observed and flow cytometry analyses showed a gradual increase in the percentage of BMSCs acquiring calcein with time. Cocultures that included BMSCs expressed significantly more type 2 collagen compared with the control.

The current study, for the first time, reported the expression of gap-junctional communication of BMSCs with two adherent cell lines of musculoskeletal system in vitro and also confirmed that incorporation of stem cells augments fibrocartilage regeneration."

couldn't get full study.

Mesenchymal stem cells derived from synovium, meniscus, anterior cruciate ligament, and articular chondrocytes share similar gene expression profiles.

"Human synovium, meniscus, intraarticular ligament, muscle, adipose tissue, and bone marrow were harvested, and colony-forming cells were analyzed. All these cells showed multipotentiality and surface markers typical of MSCs. Gene profiles of intraarticular tissue MSCs and chondrocytes were closer to each other than those of extraarticular tissues MSCs. Among three characteristic genes specific for intraarticular tissue MSCs, we focused on proline arginine-rich end leucine-rich repeat protein (PRELP). Higher expression of PRELP was confirmed in chondrocytes and intraarticular tissue MSCs among three elderly and three young donors. Synovium MSCs stably expressed PRELP, contrarily, bone marrow MSCs increased PRELP expression during in vitro chondrogenesis. In conclusion, MSCs could be isolated from various intraarticular tissues including meniscus and ligament, gene expression profiles of intraarticular tissue MSCs closely resembled each other, and the higher expression of PRELP was characteristic of intraarticular tissue MSCs."

Pellets from the synovium and bone marrow were over 1mg indicative of chondrogenic potential.

PRELP was expressed higher in chondrocytes and intra-articular MSCs than other tissues.

"OGN{down} and ECRG4 expressions were higher in synovium MSCs than in bone marrow MSCs"

DSP is expressed lowever in cartilage than bone marrow and is downregulated in LSJL.

"Synovium MSCs expressed PRELP stably during the chondrogenesis, contrarily, bone marrow MSCs increased PRELP expression along with the chondrocyte differentiation"

Expression profiles of two types of human knee-joint cartilage.

"We have performed a comprehensive analysis of gene-expression profiles in human articular cartilage (hyaline cartilage) and meniscus (fibrocartilage) by means of a cDNA microarray consisting of 23,040 human genes. Comparing the profiles of the two types of cartilage with those of 29 other normal human tissues identified 24 genes that were specifically expressed in both cartilaginous tissues; these genes might be involved in maintaining phenotypes common to cartilage. We also compared the cartilage profiles with gene expression in human mesenchymal stem cells (hMSC), and detected 22 genes that were differentially expressed in cells representing the two cartilaginous lineages, 11 specific to each type, which could serve as markers for predicting the direction of chondrocyte differentiation."

Genes expressed both in hyaline and fibrocartilage also expressed in LSJL:

Sox9(Sox9 was also expressed in non-cartilagenous samples however)

MMP3

Genes only in hyaline cartilage and not fibrocartilage versus MSCs also upregulated in LSJL:

Col9a1

Genes only in fibrocartilage:

HARS{down}

Nap1l4{down}

Lox

Lmna

Col1a1

IGF2 is a marker gene for cartilage over fibrocartilage.

CTRP1

Also known as C1QTNF1 which LSJL does upregulate.

A novel adipokine C1q/TNF-related protein 1 (CTRP1) regulates chondrocyte proliferation and maturation through the ERK1/2 signaling pathway.

"CTRP1, a paralog of adiponectin, is a member of the C1q and tumor necrosis factor (TNF)-related protein (CTRP) superfamily. It is expressed at high levels in adipose tissue and has recently emerged as a novel adipokine.  CTRP1 [has a role] in chondrocyte proliferation and maturation [in] a mouse chondrocytic cell line, N1511. The CTRP1 protein was strongly expressed and predominantly distributed in the reserve and proliferative chondrocytes in the fetal growth plate and its mRNA decreased during the maturation of N1511 chondrocytes. Recombinant CTRP1 promoted proliferation of immature proliferating N1511 chondrocytes in a dose-dependent manner, whereas it inhibited maturation of maturing N1511 chondrocytes. The stimulatory effect of CTRP1 on chondrocyte proliferation was associated with activation of the extracellular signal-regulated kinases 1/2 (ERK1/2) signaling pathway. The inhibitory effect of CTRP1 on chondrocyte maturation is associated with suppression of the ERK1/2 pathway."

So CTRP1 could play a role in the LSJL height increasing effect.

"CTRP1 [is] expressed in vascular wall tissues and [inhibits] collagen-induced platelet aggregation by blocking the binding of von Willebrand factor to collagen"

"Staining [for CTRP1] was observed in articular chondrocytes but was restricted to the superficial and middle zone"

"increased ERK1/2 phosphorylation in [immature chondrocyte] cells stimulated with 10 μg/ml of CTRP1 after 15 min, with the increase peaking after 30 min of stimulation and decreasing after 1 h"

"CTRP1 had no effect on the activities of JNK1/2, p38 MAPK, and Akt, and none of their phosphorylated forms was detected"

"Inhibition of the ERK1/2 pathway by pretreatment with U0126 led to a block of CTRP1-stimulated proliferation of immature proliferative chondrocytes"

"[Due to CTRP1] the expression levels of phosphorylated ERK1/2 gradually decreased within 5 min, and this inhibitory effect lasted for at least 1 h in maturing chondrocytes"

"The expression of mRNA for Col2al and aggrecan in the U0126-treated[the ERK 1/2 inhibitor] [in addition to CTRP1] maturing N1511 chondrocytes was reduced to 52% and 43%"

"When immature proliferating N1511 chondrocytes were induced to mature by BMP-2 and insulin treatment, there was a significant decrease in levels of CTRP1 mRNA."

"CTRP3 [has a stimulatory effect] on the proliferation of immature proliferating chondrocytes in N1511 cell cultures"<-CTRP3 is upregulated over 6 fold in LSJL as C1qtnf3.

"Unlike the immature proliferating chondrocytes, the maturing chondrocytes have a high basal level of activated ERK1/2, and the high basal phosphorylation of ERK1/2 is suppressed following CTRP1 treatment."<-So ERK1/2-p is biphasic on growth.

"During chondrocyte maturation, high levels of Col2a1 and aggrecan mRNA require persistently high basal activation of the ERK1/2 pathway"

"transforming growth factor-β (TGF-β) increased ERK1/2 phosphorylation in undifferentiated ATDC5 chondrocytes but decreased phosphorylated ERK1/2 in differentiated ATDC5 cells."

How to Grow Taller If You're a Teenage Girl

Nobody wants to be told how they should look like.  The issue with height increase in teenage girls is that many typical height gaining compounds or activities are considered mainly like Testosterone or IGF-1.  While you may hear that many men prefer short, voluptuous women ultimately you have to look good for yourself.  You have to decide what chemicals or methods to apply on your body in order to grow taller.

Ways to Grow Taller that don't Involve Looking Manly

The best and least controversial way to increase height growth during puberty is by anti-oxidants.  Anti-oxidants essentially protect your growth plate cells from damage.  Nothing manly about that.  Anti-Oxidants:

Green Tea: Tazo China Green Tips Green Tea, 20-Count Tea Bags (Pack of 6)
Vitamin A, C, E:  Spring Valley Antioxidant
Melatonin:  Source Naturals Melatonin 2.5mg, Peppermint, 240 Tablets
Selenium: Selenium 200mcg 100 caps
Zinc:  Spring Valley Zinc
Acai Berry: Life Smart Labs, 1300 mg Acai Berry Juice Extreem TM New Stronger Potency HIGH 
Astragalus Membranaceous: Astragalus ( Astragalus membranaceus ) 1 Oz Herb Pharm

Then there are other protectors that protect growth plate DNA from damage.

Folic Acid: Nature Made Folic Acid 400mcg, 250 Tablets (Pack of 3)
The B Vitamins: Nature Made Super Vitamin B-Complex with Vitamin C - 300 Tablets

You can get many of these compounds from foods.  Also, sleep as well helps you fight off free radicals by learning your metabolic rate.

Now, here are some chemical methods that don't involve Testosterone but it's very hard to just stimulate the bone.  These two compounds stimulate the PI3K pathway and will increase cellular proliferation in all cells including growth plate cells:

Puerarin: Planetary Herbals Full Spectrum Kudzu, 750 mg, Tablets , 240 tablets
Leptin: Nature's Vision New Balance w/ LEPTIN Homeopathic Liquid - 2 fl oz

Now, there's mechanical methods which includes Lateral Synovial Joint Loading.  Lateral Synovial Joint Loading applies lateral loads directly to the long bones thus, you can control which body parts you want to get better.  If you don't want to build muscle, you can stick with the clamp only.  Since you are in development, you don't have to use as much pressure as an adult.

There's also LIPUS which increases cellular proliferation.  You can apply it directly to the growth plates.  Here's an ultrasound machine but note that the studies on Ultrasound have been done on the $3,000 version: ReliaMed Portable Ultrasound.

All of these methods will allow you to grow taller and stay "girly".  If that's not enough then there are other methods that are less "feminine".

Does dieting stunt growth?

In order, to truly understand the effect of diet on height growth, we have to understand the effects of caloric deficit and surplus on chondrocytes and osteoblasts(other cell types may affect height growth as well, not to mention endocrinological and nutritional effects).  We have to understand if caloric surplus and deficit alone have an effect on the cells that can increase height and not just an indirect effect via say a nutritional deficiency which can be alleviated by supplements.  And then there's the whole issue of growth rate versus final adult height.

Before we learned that Leptin and IGF-1 are downregulated during fasting and upregulated during catch-up growth.  Leptin and IGF-1 stimulate the PI3K pathways that stimulate cellular proliferation.  Levels of GHR and IGF-1R went down as well during fasting, however, the mRNA levels remained the same so they should be able to recover efficiently during catch up growth.  Other chondrocytes should operate similarly including articular chondrocytes(which can undergo endochondral ossification during osteoarthritis) and hydrostatic pressure induced chondrocytes. 


Leptin reverses the inhibitory effect of caloric restriction on longitudinal growth.

"Caloric imbalance, particularly in critical periods of growth and development, is often the underlying cause of growth abnormalities. Serum levels of leptin are elevated in obesity and are low in malnutrition and malabsorption. The aim of the present study was to determine whether leptin integrates energy levels and growth in vivo even in the presence of caloric restriction. In the first part of the study, mice were divided into three groups. Two groups were fed ad libitum[when hungry] and received leptin or vehicle only, and the third group was pair-fed with the group injected with leptin to dissociate leptin's effect on growth from its effect on food consumption. Mice given leptin had a significantly greater tibial length than untreated pair-fed animals and a similar tibial length as control mice fed ad libitum despite their lower weight. In addition, leptin significantly increased the overall size of the epiphyseal growth plate by 11%. On immunohistochemistry and in situ hybridization studies, leptin stimulated both the proliferation and differentiation of tibial growth plate chondrocytes without affecting the overall organization of the plate. There was also a marked increase in the expression and level of IGF-IR. In the second part of the study, two groups of mice were fed only 60% of their normal chow; one was injected with leptin, and the other was injected with vehicle alone. Caloric deprivation by itself reduced serum levels of IGF-I by 70% and the length of the tibia by 5%. Leptin treatment corrected the fasting-induced growth deficiency, but further reduced the level of serum IGF-I. These results indicate that leptin stimulates growth even in the presence of caloric restriction independently of peripheral IGF-I." 


Leptin is produced by adipocytes which is related to caloric surplus and restriction.  However, you could have a lot of body fat and be under caloric restriction for example. You can take Leptin as a supplement.  This supplement for example contains Leptin(not sure if there are better deals) and it also contains green tea (which inhibits PGE2 and COX2) which may have beneficial effects on height: Lepti-Trim Night Time Formula (16 oz). 

So, caloric restriction may have height lowering effects as a result of lowering Leptin and IGF-1 levels but caloric restriction may not independently have any height decreasing effects.  The amount of Leptin receptors change with age so altering the number of Leptin receptors may affect height growth.


Age-related variations of leptin receptor expression in the growth plate of spine and limb: gender- and region-specific changes.

"Leptin is a potent growth-stimulating factor of bone. The effects of leptin on bone growth differ significantly between axial and appendicular regions. Gender differences of leptin function have also been suggested in normal pubertal development. To explore the mechanisms underlying these effects, we investigated the spatial and temporal expressions of the active form of the leptin receptor (Ob-Rb) in the tibial and spinal growth plates of the female and male rats during postnatal development. The 1-, 4-, 7-, 12- and 16-week age stages are representative for early life, puberty and early adulthood after puberty, respectively. Quantitative real-time PCR was used for Ob-Rb mRNA examination and comparison. The spatial location of Ob-Rb was determined by immunohistochemical analysis. There were gender- and region-specific differences in Ob-Rb mRNA expression in the growth plate. Mainly cytoplasm staining of Ob-Rb immunoreactivity was observed in the spinal and tibial growth plate chondrocytes of both genders. Spatial differences of region- and gender-related Ob-Rb expression were not observed. Ob-Rb immunoreactivity was detected in the resting, proliferative and prehypertrophic chondrocytes in early life stage and during puberty. After puberty, staining was mainly located in the late proliferative and hypertrophic chondrocytes[So new chondrocytes no longer have leptin receptors?]. The results of Ob-Rb HSCORE analysis were similar to those obtained from quantitative real-time PCR. Our study indicated direct effects on the chondrocytes of the growth plate in different development stages. The region-specific expression patterns of Ob-Rb gene might be one possible reason for contrasting phenotypes in limb and spine. Different Ob-Rb expression patterns might partly contribute to age- and gender- related differences in trabecular bone mass."

Why would new chondrocytes lose leptin receptors over time?  Maybe it has to do with methylation status or telomere length?  Newer cells may be undermethylated.  Maybe the Ob-Rb status serves to regulate chondrocyte proliferative capacity.

"By affecting the proliferation, hypertrophy and calcification of chondrocytes through Ob-Rb, leptin has a direct effect on longitudinal growth. The balance among chondrocytes of different zone is crucial for bone metabolic regulation in the growth plate. The rate of the proliferative chondrocytes and accelerated or delayed differentiation could lead to abnormal longitudinal growth of bone. The regulation process is controlled by various growth factors/hormones via their receptors. In the present study, Ob-Rb immunostaining was mainly revealed in the cytoplasm of the chondrocytes in the tibial and spinal growth plates. Low staining in nuclei of chondrocytes was also detected. These results indicate the main target of leptin in the chondrocytes at different stages and layers of the growth plate."<-The number of Leptin receptors is important.  How do we increase the number of those receptors?


Determinants of height in adolescent girls with anorexia nervosa.

"Anorexia nervosa, a condition characterized by marked caloric restriction and low insulin like growth factor-1 levels, would be expected to cause short stature. However, this disorder is also associated with hypogonadotropic hypogonadism[basically a deficiency in sex hormones] and high growth hormone levels. Delays in growth-plate closure from associated hypogonadism may result in a longer period of time available for statural growth with protective effects on stature[growth plates don't close, senescence followed by ossification]. In addition, growth hormone may have direct effects on the growth plate independent of insulin-like growth factor 1 to increase statural growth.
To determine the impact of undernutrition, hypogonadism, and acquired growth hormone resistance on height in adolescents with anorexia nervosa (aged 12-18 years), we examined 208 girls: 110 with anorexia nervosa and 98 controls of comparable chronological age. Sixty-three girls with anorexia nervosa and 79 controls were followed prospectively over 1 year. Mean duration of illness was 11.6 +/- 13.2 months. In a subset, overnight growth hormone sampling was performed every 30 minutes for 12 hours, and fasting insulin-like growth factor 1 levels were obtained.
The difference between height and target height and between predicted adult height and target height did not differ between the groups, indicating preservation of height potential. The groups had comparable bone age, but bone age was lower than chronological age in girls with anorexia nervosa. Girls with anorexia nervosa had lower insulin-like growth factor 1 levels and higher nadir growth hormone levels than those of controls. Nadir growth hormone levels predicted height SD scores and predicted adult-height SD scores in controls but not in the girls with anorexia nervosa. In girls with anorexia nervosa, insulin-like growth factor 1 and duration of illness predicted height measures. Height SD scores of <0 were more likely after 32 months of illness and at insulin-like growth factor 1 levels of <134 ng/mL. Delayed baseline bone age predicted subsequent increases in height SD scores in immature girls with anorexia nervosa.
Our data suggest that preservation of height potential in this cohort of girls with anorexia nervosa may be a consequence of delayed bone age. Hypogonadism may negate the deleterious effects of undernutrition on stature by allowing for a longer duration of growth."

So, Height is conserved during Anorexia Nervosa.  Now some may point out that the AN girls produced less Estrogen, however Estrogen needs to be kept in a certain range for optimal height growth although low levels of estrogen are much less detrimental to height growth than high levels.  Girls are more likely to be above this Estrogen range than males.  Thus the Anorexia may have knocked them out of the high end of Estrogen.

However, it is more likely that growth is conserved during periods of under nutrition.

So, basically the benefits of caloric restriction and surplus are mainly affected by Leptin and IGF-1.  Leptin of which is available as a supplement.

Height Increase by your Extracellular Matrix

It was found that cell space affected cellular senescence.  Growth plates tend to be tightly packed against each other without sufficient Extracellular Matrix.  Dynamic compression of chondrocytes increases ECM matrix synthesis.  Although the amount of ECM decreases the response of cells to stimuli such as vibration.  Smoking in high doses decreases ECM synthesis(in low doses it may increase it).  Chondroitin and Glucosamine are two supplements involved in the ECM as is Hyaluronic Acid which is likely the most effective.  Also, inhibiting Smads 1/5/8 has been shown to enhance height growth whereas Smads 2/3 increase height(as a result of increasing Collagen Type II production).It's long been thought that chondrocyte hypertrophy plays the largest role in growing taller but what if the extracellular matrix is involved as well?  The ECM serves as an elastic substance that allows the bone to stretch and thus for us to increase our heights. 

Human growth plate contains aggrecan fragments that can be generated by m-calpain. 

"Growth plate is a cartilaginous structure responsible for longitudinal growth and calcification of long bones. Aggrecan is initially expressed within the growth plate but is lost during the course of calcification[Aggrecan is a member of the Chondroitin Sulfate Family]. Calcium-activated proteinases are believed to play a primary role in aggrecanolysis[lysis is a form of cell death]. In this study, m-calpain was shown to be expressed in a limited area of the growth cartilage at the lower hypertrophic zone. Confocal immunostaining demonstrated colocalization of m-calpain and the aggrecan product within the lower hypertrophic chondrocytes and in limited region of the pericellular matrix. Immunoblotting analysis identified anti-VPGVA-positive aggrecan product within the dissociative fractions of A1D1-A1D6 (densities 1.65, 1.56, 1.52, 1.47, 1.41, and 1.37 g/cm(3), respectively). These findings indicated that limited expression of m-calpain is colocalized with the appearance of calpain-related aggrecan products at the sites of aggrecanolysis and calcification, and suggested an important role of m-calpain in regulation of the growth plate process. The G1-G2-containing fragment of aggrecan remaining within the extracellular matrix (ECM) of the growth plate may contribute to the mechanical properties of the growth plate between the chondrocyte cell layers until bony replacement takes place."

The loss of aggrecan is important to chondrogenic differentiation into osteogenic cells. By increasing aggrecan levels it may be possible to elongate the period before osteogenic differentiation and thus grow taller for a longer period of time.

Sox9 is important in ECM formation. 

Identification of SOX9 interaction sites in the genome of chondrocytes.

"The transcription factor SOX9 is completely needed for chondrogenic differentiation and cartilage formation acting as a "master switch" in this differentiation. Heterozygous mutations in SOX9 cause campomelic dysplasia, a severe skeletal dysmorphology syndrome in humans characterized by a generalized hypoplasia of endochondral bones. To obtain insights into the logic used by SOX9 to control a network of target genes in chondrocytes, we performed a ChIP-on-chip experiment using SOX9 antibodies.

The ChIP DNA was hybridized to a microarray, which covered 80 genes, many of which are involved in chondrocyte differentiation. Hybridization peaks were detected in a series of cartilage extracellular matrix (ECM) genes including Col2a1, Col11a2, Aggrecan and Cdrap as well as in genes for specific transcription factors and signaling molecules. Our results also showed SOX9 interaction sites in genes that code for proteins that enhance the transcriptional activity of SOX9. Interestingly, a strong SOX9 signal was also observed in genes such as Col1a1 and Osx, whose expression is strongly down regulated in chondrocytes but is high in osteoblasts. In the Col2a1 gene, in addition to an interaction site on a previously identified enhancer in intron 1, another strong interaction site was seen in intron 6. This site is free of nucleosomes specifically in chondrocytes suggesting an important role of this site on Col2a1 transcription regulation by SOX9."

"Chondrogenesis is also controlled by a complex interplay of signaling molecules among which some target either the expression or the activity of SOX9. Whereas IL-1 and TNF α inhibit its expression, FGF signaling increases its expression and its activity; Wnt/β-catenin also inhibits its activity and expression, whereas PTHrP increases its activity."

Calcium has been shown to inhibit PTH and increase Beta-Catenin but calcium deficiency decreases height.  Beta Catenin also increases chondrocyte hypertrophy.  The negative height gaining aspects of Calcium must be outweighed by the positive.  CARM1 which disrupts the interaction of Sox9 with Beta-Catenin has been shown to result in you becoming taller.  So you need a precise blend of compounds to optimally grow taller.

LIPUS increases ECM related genes. 

Low-intensity pulsed ultrasound stimulates cell proliferation, proteoglycan synthesis and expression of growth factor-related genes in human nucleus pulposus cell line.

"Low-intensity pulsed ultrasound (LIPUS) stimulation has been shown to effect differentiation and activation of human chondrocytes. A study involving stimulation of rabbit disc cells with LIPUS revealed upregulation of cell proliferation and proteoglycan (PG) synthesis. However, the effect of LIPUS on human nucleus pulposus cells has not been investigated. In the present study, therefore, we investigated whether LIPUS stimulation of a human nucleus pulposus cell line[the cells in the intervertebral disc that affect disc height, so you should also be able to grow taller that way but the affects on matrix synthesis should be applicable to other chondrocyte matrix production namely growth plates] (HNPSV-1) exerted a positive effect on cellular activity. HNPSV-1 cells were encapsulated in 1.2% sodium alginate solution at 1x10(5) cells/ml and cultured at 10 beads/well in 6-well plates. The cells were stimulated for 20 min each day using a LIPUS generator, and the effects of LIPUS were evaluated by measuring DNA and PG synthesis. Furthermore, mRNA expression was analyzed by cDNA microarray using total RNA extracted from the cultured cells. Our study revealed no significant difference in cell proliferation between the control and the ultrasound treated groups. However, PG production was significantly upregulated in HNPSV cells stimulated at intensities of 15, 30, 60, and 120 mW/cm(2) compared with the control. The results of cDNA array showed that LIPUS significantly stimulated the gene expression of growth factors and their receptors (BMP2, FGF7, TGFbetaR1 EGFRF1, VEGF). These findings suggest that LIPUS stimulation upregulates PG production in human nucleus pulposus cells by the enhancement of several matrix-related genes including growth factor-related genes."

So LIPUS may be able to increase height by increasing ECM in growth plate cells and by directly increasing the height of the intervertebral discs.

The periosteum may be important in ECM function as well... 

Development of bone and cartilage in tissue-engineered human middle phalanx models.

"Human middle phalanges were tissue-engineered with midshaft scaffolds of poly(L-lactide-epsilon-caprolactone) [P(LA-CL)], hydroxyapatite-P(LA-CL), or beta-tricalcium phosphate-P(LA-CL) and end plate scaffolds of bovine chondrocyte-seeded polyglycolic acid. Midshafts were either wrapped with bovine periosteum or left uncovered. Constructs implanted in nude mice for up to 20 weeks were examined for cartilage and bone development as well as gene expression and protein secretion, which are important in extracellular matrix (ECM) formation and mineralization. Harvested 10- and 20-week constructs without periosteum maintained end plate cartilage but no growth plate formation. They also consisted of chondrocytes secreting type II collagen and proteoglycan, and they were composed of midshaft regions devoid of bone. In all periosteum-wrapped constructs at like times, end plate scaffolds held chondrocytes elaborating type II collagen and proteoglycan and cartilage growth plates resembling normal tissue. Chondrocyte gene expression of type II collagen, aggrecan, and bone sialoprotein varied depending on midshaft composition, presence of periosteum, and length of implantation time. Periosteum produced additional cells, ECM, and mineral formation within the different midshaft scaffolds. Periosteum thus induces midshaft development and mediates chondrocyte gene expression and growth plate formation in cartilage regions of phalanges. This work is important for understanding developmental principles of tissue-engineered phalanges and by extension those of normal growth plate cartilage and bone."

So things that are anabolic to the periosteum may have additional affects on ECM and cartilagenous growth plates in such a way as to increase height.  Although, sprinters who generate a lot of shear strain on the periosteum were not found to be taller than non-sprinters(in fact they were a little bit shorter).  But this could be due to the fact that perhaps shorter individuals prefer running.

The Extracellular Matrix also provides cues for height growth, remember that Type I Collagen is essential for chondrogenic differentiation of stem cells(Type I Collagen is a huge part of bone tissue):

Mesenchymal Stem Cells Sense Three Dimensional Type I Collagen through Discoidin Domain Receptor 1.

"The extracellular matrix provides structural and organizational cues for tissue development and defines and maintains cellular phenotype during cell fate determination. Multipotent mesenchymal stem cells use this matrix to tightly regulate the balance between their differentiation potential and self-renewal in the native niche. When understood, the mechanisms that govern cell-matrix crosstalk during differentiation will allow for efficient engineering of natural and synthetic matrices to specifically direct and maintain stem cell phenotype. This work identifies the discoidin domain receptor 1 (DDR1), a collagen activated receptor tyrosine kinase, as a potential link through which stem cells sense and respond to the 3D organization of their extracellular matrix microenvironment. DDR1 is dependent upon both the structure and proteolytic state of its collagen ligand and is specifically expressed and localized in three dimensional type I collagen culture. Inhibition of DDR1 expression results in decreased osteogenic potential, increased cell spreading, stress fiber formation and ERK1/2 phosphorylation. Additionally, loss of DDR1 activity alters the cell-mediated organization of the naïve type I collagen matrix. Taken together, these results demonstrate a role for DDR1 in the stem cell response to and interaction with three dimensional type I collagen. Dynamic changes in cell shape in 3D culture and the tuning of the local ECM microstructure, directs crosstalk between DDR1 and two dimensional mechanisms of osteogenesis that can alter their traditional roles."

So proper DDR1 signaling is essential for height growth and for LSJL.

"Changes in cell shape, cytoskeletal tension and tissue geometry influence cell fate decisions[lateral joint loading influences the last two by tension against the actin cytoskeleton and bone deformation, it possibly influences cell shape by dynamic compression of chondrocytes]. Cell shape controls two dimensional (2D) cell behavior through alteration of internal cytoskeletal tension"

The spatial properties of the ECM also play a role in cell fate.  This is a problem for height growth post puberty but not an insurmountable one.  Of course, this is the factors that affect cells on two planes whereas within the bone there are three.

"Cell-mediated reorganization of the ECM requires adhesion, secretion, degradation and assembly of the proteins within that matrix. DDR1 and integrins are responsible for the upregulation of matrix metalloproteinases (MMPs) in response to collagen adhesion that mediate the cellular remodeling response."

DDR upregulates MMPs.  So, even if post-fusion there is no cartilagenous template.  DDR1 may allow for the formation of new cartilage canals to form new ones.

"Geometric control of cell shape by changing ECM dynamics alters intracellular distances. Spatial restriction, in this way, can force crosstalk between pathways that would not cross in a planar cell."

Cellular Compression alters intracellular distances that changes cross talk dynamics.  Lateral Joint Loading may alter both intracellular and intercellular distances.

ECM Stiffness Primes the TGFβ Pathway to Promote Chondrocyte Differentiation.

"Cells encounter physical cues such as extracellular matrix (ECM) stiffness in a microenvironment replete with biochemical cues.  Here we investigate mechanisms by which chondrocytes generate an integrated response to ECM stiffness and TGFβ, a potent agonist of chondrocyte differentiation. Primary murine chondrocytes and ATDC5 cells grown on 0.5 MPa substrates deposit more proteoglycan and express more Sox9, Col2α1, and aggrecan mRNA relative to cells exposed to substrates of any other stiffness. The chondroinductive effect of this discrete stiffness, which falls within the range reported for articular cartilage, requires the stiffness-sensitive induction of TGFβ1. Smad3 phosphorylation, nuclear localization, and transcriptional activity are specifically increased in cells grown on 0.5 MPa substrates. ECM stiffness also primes cells for a synergistic response, such that the combination of ECM stiffness and exogenous TGFβ induce chondrocyte gene expression more robustly than either cue alone through a p38 MAPK-dependent mechanism. In this way, the ECM stiffness primes the TGFβ pathway to efficiently promote chondrocyte differentiation. This work reveals novel mechanisms by which cells integrate physical and biochemical cues to exert a coordinated response to their unique cellular microenvironment."

What's the stiffness of the adult epiphyseal bone marrow?

Growth and differentiation of pre-chondrogenic cells on bioactive self-assembled peptide nanofibers.

"Sulfated glycosaminoglycan molecules are vital constituents of both developing and mature cartilage extracellular matrix. The interplay between regulator proteins and glycosaminoglycan molecules has an essential role in coordinating differentiation, expansion and patterning during cartilage development. We exploited the functional role of extracellular matrix on chondrogenic differentiation by imitating extracellular matrix both chemically by imparting functional groups of native glycosaminoglycans, and structurally through peptide nanofiber network. For this purpose, sulfonate, carboxylate and hydroxyl groups were incorporated on self-assembled peptide nanofibers. When ATDC5 cells were cultured on functional peptide nanofibers, they rapidly aggregated in insulin-free medium and formed cartilage-like nodules and deposited sulfated glycosaminoglycans. Collagen II and aggrecan gene expressions were significantly enhanced which indicated the remarkable bioactive role of this nanofiber system on chondrogenic differentiation. Glycosaminoglycan mimetic peptide nanofiber system provides a promising platform for cartilage regeneration."

"Perlecan, a heparan sulfate proteoglycan, functions as a growth factor reservoir, thereby increasing local concentration of growth factors"  It also helps trigger chondrocyte differentiation.

"proteoglycans are highly negatively charged biomacromolecules in cartilage extracellular matrix. The carboxylate and sulfate groups on proteoglycans provide fixed negative charge to extracellular matrix and each proteoglycan-associated negative charge requires a mobile counter ion to maintain tissue electroneutrality."

"The mobile counter ions (e.g. Na+) coming from outside of the tissue result in drawing of water into the tissue and high swelling pressure."

"NF 4[the most chondrogenic group] was composed of mainly E-PA and SO3-PA that bring carboxylate, hydroxyl and sulfonate groups together at proper ratio."

So how to grow taller via the ECM:

• Periosteum 
• LIPUS 
• SOX9
• ECM may help control the differentiation of stem cells and thus may help them to undergo a chondrogenic lineage, DDR1 helps the cells interact with the ECM
• Supplements that increase ECM formation or decrease ECM degradation(some degradation is needed for proper modeling)

Grow Taller with Anti-oxidants

This blog has spoken about Reactive Oxygen Species(free radicals) causing DNA damage before with Anti-Oxidants such as Astragalus, Acai Berry, Vitamin E, Phlomis Umbrosa, Melatonin, and Vitamin C all helping to fight off this damage.  As your activities increase more and more you produce more free radicals and thus you need more anti-oxidants.  One way to counteract this is via sleep which reduces the metabolic rate and gives the body the ability to mop up free radicals while not under so much pressure.  The more you increase your activity the more anti-oxidants or sleep you need.  Why are anti-oxidants so important for height increase?


Cell Contact Accelerates Replicative Senescence of Human Mesenchymal Stem Cells Independent of Telomere Shortening and p53 Activation: Roles of Ras and Oxidative Stress.


"Before transplantation, in vitro culture-expansion of MSCs is necessary to get desired cell number. We observed that cell contact accelerated replicative senescence during such process. To confirm the finding as well as to investigate the underlying mechanisms, we cultured both human bone marrow- and umbilical cord blood-derived MSCs under non-contact culture (subculture performed at 60-70% of confluence), or contact culture (cell passage performed at 100% of confluence). It was found that MSCs reached cellular senescence earlier in contact culture, and the doubling time was significantly prolonged. Marked increase of senescence-associated beta-galactosidase positive staining was also observed as a result of cell contact. Cell cycle analysis revealed increased frequency of cell cycle arrest after contact culture. It was noted, however, that the telomere length was not altered during contact-induced acceleration of senescence. Moreover, cell cycle checkpoint regulator P53 expression was not affected by cell contact. Marked increase in intracellular reactive oxygen species (ROS) and a concomitant decrease in the activities of anti-oxidative enzymes were also observed during contact-induced senescence. Importantly, increased p16INK4a following Ras up-regulation was found after contact culture. Taken together, cell contact induced accelerated senescence of MSCs, which is telomere shortening- and p53-independent. ROS accumulation due to defective ROS clearance function together with Ras and p16INK4a up-regulation play an important role in contact-induced senescence of MSCs. Over-confluence should therefore be avoided during in vitro culture-expansion of MSCs in order to maintain their qualities."


The inability to clear out reactive oxygen species resulted in cellular senescence.  If you look at HGH growth plates(the pictures are on page 4), you can see that growth plates without HGH are densely packed together whereas without HGH, the number of cells are about the same but they are further spread apart.  Therefore, increasing the amount of Extracellular Matrix within the growth plate may be a way to increase height be reducing cellular contact and therefore slowing down senescence.  Note that LSJL increases ECM.  Also, things like Hyaluronic Acid inhibits MMP-13 which degrades Extracellular Matrix.

So three methods of counteracting cell-contact based growth plate chondrocyte senescence:

1) Beta-Galactosidase inhibition
2)Decreasing cell contact by increasing ECM
3) Increasing the number of anti-oxidants to compensate for the cell contact inhibiting the ability for growth plate chondrocytes to fight off free radicals.

But Reactive Oxygen Species may also be important for growth...


Reactive oxygen species generated by NADPH oxidase 2 and 4 are required for chondrogenic differentiation.


"Although generation of reactive oxygen species (ROS) by NADPH oxidases (Nox) is thought to be important for signal transduction in nonphagocytic cells, little is known of the role ROS plays in chondrogenesis. We therefore examined the possible contribution of ROS generation to chondrogenesis using both ATDC5 cells and primary chondrocytes derived from mouse embryos. The intracellular level of ROS was increased during the differentiation process, which was then blocked by treatment with the ROS scavenger N-acetylcysteine(NAC is a dietary supplement). Expression of Nox1 and Nox2 was increased upon differentiation of ATDC5 cells and primary mouse chondrocytes, whereas that of Nox4, which was relatively high initially, was decreased gradually during chondrogenesis. In developing limb, Nox1 {LSJL upregulates Nox1} and Nox2 were highly expressed in prehypertrophic and hypertrophic chondrocytes. However, Nox4 was highly expressed in proliferating chondrocytes and prehypertrophic chondrocytes. Depletion of Nox2 or Nox4 expression by RNA interference blocked both ROS generation and differentiation of ATDC5 cells, whereas depletion of Nox1 had no such effect. We also found that ATDC5 cells depleted of Nox2 or Nox4 underwent apoptosis. Further, inhibition of Akt phosphorylation along with subsequent activation of ERK was observed in the cells. Finally, depletion of Nox2 or Nox4 inhibited the accumulation of proteoglycan in primary chondrocytes. Taken together, our data suggest that ROS generated by Nox2 or Nox4 are essential for survival and differentiation in the early stage of chondrogenesis."

If you look at the IGF-1 growth plates, you can see that the chondrocytes are more densely packed at all stages of differentiation. Maybe densely packing of chondrocytes stops the scavenging of Nox1, Nox2, and Nox4 and enables the chondrocytes to undergo further differentiation.

The solution is to find anti-oxidants that don't target Nox1, Nox2, and Nox4. So, don't take NAC if you want to grow taller. Apocynin inhibits the Nox's also.

Now the NADPH Oxidases are required for Chondrogenic Differentiation but as long as they are there we don't know if extra NOX's help increase height.  Probably not.  So as long as you aren't completely inhibiting the NOX's you should be fine in terms of growing taller.  Thus, focus on increasing the amount of Extracellular Matrix to decrease cell-to-cell contact to reduce growth plate senescence and grow taller.

Rogaine can make you taller?

Minoxidil is most commonly known as Rogaine(Minoxidil-5% Extra Strength Hair Regrowth for Men, 6 Month Supply-Not sure how effectively this could be absorbed by our target cells like this within the bone).  A recent post over at the grow tall forum stated that rogaine can be involved in a type of acromegaly called psuedoacromegaly. 

Pseudoacromegaly induced by the long-term use of minoxidil. 

"Acromegaly is an endocrine disorder caused by chronic excessive growth hormone secretion from the anterior pituitary gland. Significant disfiguring changes occur as a result of bone, cartilage, and soft tissue hypertrophy, including the thickening of the skin, coarsening of facial features, and cutis verticis gyrata{thickening of the scalp, the scalp contains a flat bone, it's like a long bone that's sideways so it's easier to increase height there}. Pseudoacromegaly, on the other hand, is the presence of similar acromegaloid features in the absence of elevated growth hormone or insulin-like growth factor levels. A patient with pseudoacromegaly resulted from the long-term use of minoxidil at an unusually high dose." 

"Bone and cartilage changes pri-marily affect the face and skull. The calvaria thickens, and the frontal sinuses enlarge, which leads toprominent supraorbital ridges. Anterior and inferior growth of the mandible results in prognathism and widely spaced teeth."

Minoxidil may increase growth by increasing the deposition of GAG molecules.

"IGF-1 and GH levels along with radiographs of the chest and long bones (femora andhumeri) were all normal"

First, Acromegaly is not necessarily caused by excessive growth hormone as the effectiveness of growth hormone is based on the number of growth hormone receptors. Also, it's only one patient so it could just be a coincidence.  Also, the patient didn't really have any effects on bones only skin but that could be because only the skin cells could absorb the rogaine.  It could help us grow taller though if minoxidil had anabolic affects on chondrocytes, osteoblasts, or stem cells.  There doesn't seem to be a whole lot of new studies on minoxidil and bone but there are relevant studies that could be key in relating minoxidil(Rogaine) and height growth. 

Use of minoxidil to demonstrate that prostacyclin is not the mediator of bone resorption stimulated by growth factors in mouse calvariae. 

"Growth factors, such as human transforming growth factor-alpha (hTGF alpha) and epidermal growth factor, as well as human tumor necrosis factor (hTNF) stimulate the resorption of bone in neonatal mouse calvariae in organ culture via a prostaglandin (PG)-mediated pathway. In response to such factors mouse calvariae produce substantial quantities of prostaglandin E2 (PGE2) and prostacyclin (PGI2). We have selectively inhibited the production of PGI2, but not PGE2, using the drug minoxidil and have measured the effects on stimulated bone resportion and arachidonic acid metabolism. The increased production of 6-keto-PGF1 alpha (6k-PGF1 alpha), the hydrolytic product of PGI2, stimulated by recombinant hTGF alpha and hTNF as well as murine epidermal growth factor was inhibited by minoxidil. There was no inhibition by minoxidil of PGE2 production. Despite essentially complete inhibition of stimulated 6k-PGF1 alpha formation, there was no inhibition of bone resorption. The possibility was investigated that growth factors and TNF enhanced enzymatic conversion of PGI2 to 6k-PGE1, which stimulates bone resorption in mouse calvariae with a potency about one fourth that of PGE2. Enzymatic conversion of PGI2 to 6k-PGE1 is inhibited by rutin. Rutin did not inhibit bone resorption stimulated by hTGF alpha or hTNF.  The cyclooxygenase product that acts as the mediator of bone resorption enhanced by growth factors and TNF in mouse calvariae is probably PGE2." 

"Even concentrations of minoxidil as high as 500 micromillimeters had no inhibitory effect on PGE2 production by mouse bone, although there was complete inhibition of 6k- PGF1a production enhanced by hTGFa"

So Minodixil inhibits PGI2.  Can inhibiting PGI2 affect height growth?  

Immunohistochemical detection of prostaglandin I2 synthase in various calcified tissue-forming cells in rat. 

"Localization of prostaglandin (PG) I2 synthase immunoreactivity was examined in demineralized sections of rat pulpal, periodontal and skeletal tissues using isn-1, a monoclonal antibody raised against the enzyme. chondrocytes were immunoreactive for PGI2 synthase suggesting that they are capable of producing PGI2. In odontoblasts and chondrocytes, the reactivity increased gradually with maturation{PGI2 could play a role is ossification}.  PGI2 [may regulate] the metabolism of various calcified tissues." 

"PGE, and PGI2, are the major metabolites produced by cultured chondrocytes"

Since chondrocytes become more sensitive to PGI2 as they mature(undergo endochondral ossification) it is logical to think that maybe PGI2 plays a role in terminal differentiation. 

Prostacyclin IP receptor up-regulates the early expression of C/EBPbeta and C/EBPdelta in preadipose cells. 

"Prostacyclin (PGI(2)) and its stable analogue carbacyclin (cPGI(2)) are known to trigger the protein kinase A pathway after binding to the cell surface IP receptor and to promote or enhance terminal differentiation of adipose precursor cells to adipose cells[PGI2 is involved in terminal differentiation albeit for adipose cells]. The early expression of C/EBPbeta and C/EBPdelta is known to be critical for adipocyte differentiation in vitro as well as in vivo. We report herein that in Ob1771 and 3T3-F442A preadipose cells, activation of the IP receptor by specific agonists (PGI(2), cPGI(2) and BMY 45778) is sufficient to up-regulate rapidly the expression of C/EBPbeta and C/EBPdelta. Cyclic AMP-elevating agents are able to substitute for IP receptor agonists, in agreement with the coupling of IP receptor to adenylate cyclase. Consistent with the fact that PGI(2) is released from preadipose cells and behaves as a paracrine/autocrine effector of adipose cell differentiation, the present results favor a key role of prostacyclin by means of the IP receptor and its intracellular signaling pathway in eliciting the critical early expression of both transcription factors." 

PGI2 is involved in the early expression of transcription factors that may promote terminal differentiation.  Again this is for adiposal cells(there are no studies linking PGI2 and chondrocytes) but it is logical to think that there may be a similar link between PGI2 and terminal differentiation in chondrocytes.   But here's a study about bone and PGI2(prostacyclin). 

Increased bone mass in adult prostacyclin-deficient mice. 

"Prostaglandins (PGs) are key regulatory factors that affect bone metabolism. Prostaglandin E(2) (PGE(2)) regulates bone resorption and bone formation. Prostacyclin (PGI(2)) is one of the major products derived from arachidonic acid by the action of cyclooxygenase and PGI(2) synthase (PGIS). Unlike PGE(2), there are few reports about the role of PGI(2) in bone regulation. We used PGIS knockout (PGIS(-/-)), PGIS heterozygous (PGIS(+)(/-)), and wild-type mice to investigate the role of PGI(2). PGIS(-/-) mice gradually displayed an increase in trabecular bone mass in adolescence. Adult PGIS(-/-) mice showed an increase in trabecular bone volume/tissue volume. PGIS(-/-) mice displayed increases in both bone formation and bone resorption parameters. Levels of serum osteocalcin and C-telopeptides were increased in adult PGIS(-/-) mice. The increased bone mass patterns were rescued in PGIS(-)/(tg) mice. Adult PGIS(-/-) mice displayed an overall increase in the levels of both bone formation and bone resorption parameters, which suggests that PGI(2) deficiency accelerates high bone turnover activity with a greater increase in bone mass in aging. PGI(2) may contribute to the maintenance of normal bone mass and micro-architecture in mice in age-dependent manner." 

Inhibiting PGI2 via Minoxidil definitely increases bone density.  It also increases serum osteocalcin(which increases bone formation) and C-telopeptides(basically collagen).  This alone doesn't seem like it would increase height but it shows that inhibiting PGI2 is anabolic. 

"Prostacyclin (PGI2), a product converted by COXs and PGI2 synthase (PGIS), up-regulates cAMP levels through the activation of a PGI2 receptor"

"Resident bone cells; osteoblasts and osteocytes, can produce both PGE2 and PGI2 under mechanical loading"

Interestingly, younger PGIS null mice had lower bone mass than controls but eventually had high higher bone mass.

The increase in bone turnover alone explains the psuedoacromegaly case.  However, it is possible that minoxidil mediated inhibition of PGI2 may help to increase height by slowing down terminal differentiation of chondrocytes.

Taller Height by inhibiting PGE2 and Cox-2?

LSJL downregulates COX2. Listed on the study as prostaglandin-endoperoxin synthase 2. According to Molecular pathways mediating mechanical signaling in bone, connexin hemi channels can be knocked down during fluid shear thus inhibiting PGE2 release.  Although it mentions also that in other cases PGE2 can be stimulated.

Alkoclar made an interesting statement via email "Enhanced cnp expression coupled with partial activin antagonism enables the fused tissue to gain elasticity and degrades pge2 and cox-2 expression,re-enabling vertical bone growth over time. Pge1 release is also enhanced through this stimuli,literally softening hardened periosteum.this process requires cnp signal accumulation in order to undo the long period it had been fused"


Activin inhibits cellular differentiation. Which would inhibit cells from differentiation into chondrocytes. CNP is a guanyl cyclase activator which is involved in the nitric oxide pathway. The end result is an increase in chondrocyte hypertrophy which makes you taller. Now I don't see how this would help the fused tissue gain elasticity but it is certainly worth investigating.


LIPUS enhances Prostaglandin E2 which is regulated by Cox-2. Shear Stress also increased PGE2 levels.

Here's a study that shows that contraindicates LSJL but like anything that's anabolic helping cancer to spread faster I don't think it's anything to worry about as LSJL itself won't cause the necessary changes to induce osteoarthritis or you want to take anti-oxidants to go against the inflammatory effects(although inflammation may be good for height growth).

Shear-induced interleukin-6 synthesis in chondrocytes: roles of E prostanoid (EP) 2 and EP3 in cAMP/protein kinase A- and PI3-K/Akt-dependent NF-kappaB activation.

"Mechanical overloading of cartilage producing hydrostatic stress, tensile strain, and fluid flow can adversely affect chondrocyte function and precipitate osteoarthritis (OA)[A couple of things, you are loading the epiphysis and not the cartilage; osteoarthritis has some anabolic properties and this shows that hydrostatic pressure and tensile strain have anabolic properties; Osteoarthritis also involves CD44 and an alteration in the OPG/RANKL gradient which isn't caused by hydrostatic pressure or tensile strain]. Application of high fluid shear stress to chondrocytes [stimulates the] release of pro-inflammatory mediators, matrix degradation, and chondrocyte apoptosis. Elevated levels of cyclooxygenase-2 (COX-2), prostaglandin (PG) E(2), and interleukin (IL)-6 have been reported in shear-activated chondrocytes in vitro[this is circumstantial evidence, correlation not equal causation]. PGE(2) positively regulates IL-6 synthesis in chondrocytes. Using the human T/C-28a2 chondrocyte cell line as a model system, we demonstrate that COX-2-derived PGE(2) signals via up-regulation of E prostanoid (EP) 2 and down-regulation of EP3 receptors to raise intracellular cAMP[cAMP is involved in the Nitric Oxide pathway], and activate protein kinase A (PKA)[PKA enhances Sox9 transcriptional activity and thus may help with height growth] and phosphatidylinositol 3-kinase (PI3-K)/Akt pathways. PKA and PI3-K/Akt transactivate the NF-kappaB p65 subunit via phosphorylation at Ser-276 and Ser-536, respectively. Binding of p65 to the IL-6 promoter elicits IL-6 synthesis in sheared chondrocytes. Selective knockdown of EP2 or ectopic expression of EP3 blocks PKA- and PI3-K/Akt-dependent p65 activation and markedly diminishes shear-induced IL-6 expression. Similar inhibitory effects on IL-6 synthesis were observed by inhibiting PKA, PI3-K, or NF-kappaB using pharmacological and/or genetic interventions."

IL-6 is involved in Osteoclast formation  Since PGE2 increases IL-6 synthesis it may increase osteoclast production giving chondrocytes more room to grow.  It seems like then if anything you'd want to enhance PGE2 and COX2 to soften tissue(Alkoclar recommends LIPUS which upregulates PGE2 and COX-2).

"PGE2 exerts its biological functions via binding to four distinct transmembrane G-protein-coupled receptors (GPCRs) termed E prostanoid (EP) 1, EP2, EP3, and EP4. Following PGE2 binding, the EP receptors activate distinct intracellular signaling pathways, which may account for the pleiotropic effects of this prostaglandin. EP1 couples to Gq protein and raises intracellular calcium. EP2 and EP4 elevate intracellular cAMP levels by activating adenylate cyclase via stimulatory G (Gs) proteins. The major signaling pathway of the EP3 receptor is the inhibition of adenylate cyclase, and thus reduction of intracellular cAMP, via inhibitory G (Gi) proteins. Human chondrocytes primed with exogenous PGE2 synthesize IL-6."<-may it's the raising of intracellular calcium that stimulates chondrogenesis.

"Application of fluid shear to T/C-28a2 chondrocytes induces p65 phosphorylation at both Ser-536 and Ser-276 without affecting total p65 protein levels"

If you do decide to inhibit PGE2 and COX2...

Inhibition of cyclooxygenase-2 expression and prostaglandin E2 production in chondrocytes by avocado soybean unsaponifiables and epigallocatechin gallate.'

"Production of type II collagen and aggrecan was verified by immunohistochemistry and Western blot. Chondrocytes were incubated with: (1) control media alone, (2) ASU (4 microg/ml; 8.3 microg/ml), (3) EGCG (4, 40, 400 ng/ml), or (4) the combination of ASU[Avocado Soybean unsaponifiables] and EGCG[epigallocatechini gallate, it's found in green tea] for 24h. Cells were next incubated with control medium alone or with IL-1beta (10 ng/ml) and TNF-alpha (1 ng/ml). COX-2 gene expression by real-time PCR analysis and NF-kappaB nuclear translocation by immunohistochemistry were performed after 1h of incubation. PGE(2) production was determined by immunoassay after 24h of incubation.
Equine chondrocytes responded to cytokine activation by up-regulated gene expression of COX-2 and increased PGE(2) production. Activation was associated with NF-kappaB translocation. Individually, ASU and EGCG marginally inhibited COX-2 expression and PGE(2) production in activated chondrocytes. In contrast, the combination of ASU and EGCG reduced COX-2 expression close to non-activated control levels and significantly inhibited PGE(2) production. These reductions were statistically greater than those of ASU or EGCG alone. The inhibition of COX-2 expression and PGE(2) production was associated with inhibition of NF-kappaB translocation."


PGE2 and COX2 have an affect on vertical body growth.  Whether it augments or hinders growth is unclear.  But Green Tea and Avocados affect PGE2 and COX2.


Prostaglandin E2 inhibits BMP signaling and delays chondrocyte maturation.


"While cyclooxygenases are important in endochondral bone formation during fracture healing, mechanisms involved in prostaglandin E2 (PGE2) regulation of chondrocyte maturation are incompletely understood. The present study was undertaken to determine if PGE2 effects on chondrocyte differentiation are related to modulation of the bone morphogenetic protein (BMP) signaling pathway. In primary murine sternal chondrocytes, PGE2 differentially regulated genes involved in differentiation. PGE2 induced type II collagen and MMP-13[Remember F-spondin which reduces height growth increased MMP-13 levels by as much as 30%], had minimal effects on alkaline phosphatase, and inhibited the expression of the maturational marker, type X collagen. In BMP-2-treated cultures, PGE2 blocked the induction of type X collagen. All four EP receptors were expressed in chondrocytes and tended to be inhibited by BMP-2 treatment. RCJ3.1C5.18 chondrocytes transfected with the protein kinase A (PKA) responsive reporter, CRE-luciferase, showed luciferase induction following exposure to PGE2, consistent with activation of PKA signaling and the presence of the EP2 and EP4 receptors. Both PGE2 and the PKA agonist, dibutyryl cAMP, blocked the induction of the BMP-responsive reporter, 12XSBE, by BMP-2 in RCJ3.1C5.18 chondrocytes. In contrast, PGE2 increased the ability of TGF-beta to activate the TGF-beta-responsive reporter, 4XSBE. Finally, PGE2 down-regulated BMP-mediated phosphorylation of Smads 1, 5, and 8 in RCJ3[Phosphorylating Smad 1/5/8 means to inhibit them and Smad 1/5/8 phosphorylation is  a way to grow taller!  PGE2 downregulates this.]1C5.18 cells and in primary murine sternal chondrocytes. Altogether, the findings show that PGE2 regulates chondrocyte maturation in part by targeting BMP/Smad signaling and suggest an important role for PGE2 in endochondral bone formation."


PGE2 downregulates Phosphorylation 1/5/8 and raises levels of MMP-13.  Therefore, PGE2 is bad for height growth.  Therefore, you can grow taller with avocados(100% Avocado Oil 8 Oz (Refined, light color)) and green tea(NOW Foods Green Tea Extract, 250 Capsules / 400mg) as they inhibit PGE2.  Does anyone know any other ways to inhibit PGE2?  So inhibiting PGE2 does help vertical body growth but I don't see how inhibiting PGE2 will enable elasticity.

However there's evidence that PGE2 enhances chondrogenesis.

NSAIDS inhibit in vitro MSC chondrogenesis but not osteogenesis: implications for mechanism of bone formation inhibition in man.

"Non-steroidal anti-inflammatory drugs (NSAIDs) may inhibit bone formation. We investigated whether the reported NSAID effect on bone is related to inhibition of bone marrow mesenchymal stem cell (MSC) proliferation and osteogenic and chondrogenic differentiation and evaluated both cyclooxygenase (COX)-1 and COX-2 specific drugs. The effects of seven COX-1 and COX-2 inhibitors on MSC proliferation and osteogenic and chondrogenic differentiation were tested using Vybrant, sodium 3'-[1-(phenylaminocarbonyl)- 3,4-tetrazolium]-bis (4-methoxy-6-nitro) benzene sulfonic acid hydrate (XTT), functional and quantitative assays of MSC differentiation. The MSC expression of COX-1 and COX-2 and prostaglandin E2 (PGE-2) levels were evaluated serially during lineage differentiation by quantitative PCR and ELISA. None of the NSAIDs at broad range of concentration (range 10(-3) to 100 μg/ml) significantly affected MSC proliferation. Surprisingly, MSC osteogenic differentiation inhibition was not evident. However, NSAIDs affected chondrogenic potential with a reduction in sulphated glycosaminoglycans (sGAG) content by 45% and 55% with diclofenac and ketorolac, respectively (P < 0.05 compared to controls). Parecoxib and meloxicam, more COX-2 specific reagents inhibited sGAG to a lesser degree, 22% and 27% respectively (P < 0.05 compared to controls). Cartilage pellet immunohistochemistry confirmed the above results. Pellet chondrogenesis was associated with increased COX-1 expression levels but not COX-2, and COX-1 specific drugs suppressed MSC PGE-2 more than COX-2 specific inhibitors. These findings suggest that NSAIDs may inhibit bone formation via blockage of MSC chondrogenic differentiation."

So non-steroidal anti-inflammatory drugs may decrease chondrogenesis thus decreasing height.

"[PGE2] can stimulate new bone formation, increase bone mass and also regulate both bone morphogenetic protein 2 (BMP-2) and BMP-7 expression"

Can Serotonin help you Grow Taller?

CH Turner was one of the load researchers behind Lateral Synovial Joint Loading who unfortunately passed away on July 16th,2010 which is likely why there hasn't been much research on LSJL since the groundbreaking study that found LSJL possible to increase limb length. CH. Turner was the main expert on bone research with Hiroki Yokota providing more of the gene expression analysis(mechanotransduction).  CH Turner's most likely successor is the scientist SJ Warden whom I'll be watching for any LSJL related studies.  CH Turner did leave us some artifacts in addition to his LSJL legacy.

One of his last papers was about Serotonin, a neurotransmitter.  Neurotransmitters have had height increase benefits before like with melatonin.  Busiprone stimulates serotonin receptors.

The emerging role of serotonin (5-hydroxytryptamine) in the skeleton and its mediation of the skeletal effects of low-density lipoprotein receptor-related protein 5 (LRP5).

"Novel molecular pathways obligatory for bone health are being rapidly identified. One pathway recently revealed involves gut-derived 5-hydroxytryptamine (5-HT) mediation of the complete skeletal effects of low-density lipoprotein receptor-related protein 5 (LRP5)[Serotonin helps to mediate some of the skeletal effects of LRP5]. Mounting evidence supports 5-HT as an important regulatory compound in bone with previous evidence demonstrating that bone cells possess functional pathways for responding to 5-HT[Bone cells have receptors for serotonin]. In addition, there is growing evidence that potentiation[potentiation means enhancement] of 5-HT signaling via inhibition of the 5-HT transporter (5-HTT) has significant skeletal effects. The later is clinically significant as the 5-HTT is a popular target of pharmaceutical agents, such as selective serotonin reuptake inhibitors (SSRIs)[So if inhibiting 5-HTT can increase height then SSRIs can increase height as well], used for the management of major depressive disorder and other affective conditions. The observation that 5-HT mediates the complete skeletal effects of LRP5 represents a significant paradigm shift from the traditional view that LRP5 located on the cell surface membrane of osteoblasts exerts direct skeletal effects via Wnt/beta-catenin signaling. This paper discusses the mounting evidence for skeletal effects of 5-HT and the ability of gut-derived 5-HT to satisfactorily explain the skeletal effects of LRP5."

Serotonin interacts with LRP5 to modulate bone cells. There are Serotonin reuptake inhibitors on the market that inhibit the reuptake of Serotonin. Note there are a lot of contradictory studies about Serotonin where some say it stimulates bone and others say it inhibits bone.

"5-HT receptors have been identified in all the major bone cell types (osteoblasts, osteocytes and osteoclasts), and stimulation of these receptors influences bone cell activities"

"Direct stimulatory effect on bone formation pathways with 5-HT increasing prostaglandin E₂ release from osteocyte-like (MLO-Y4) cells and enhancing proliferation of MC3T3-E1 cells and primary human osteoblasts"

"Cells (RAW264.7 and human peripheral blood mononuclear cells stimulated with RANKL) treated with 5-HT (0.01–50 μM) or a SSRI (fluoxetine; 0.001–10 μM) increased their differentiation into osteoclast-like cells and increased their bone-resorption activity"

"Binding of Wnt to its LRP5 and frizzled co-receptors inactivates glycogen synthase kinase-3β (GSK-3β) allowing for cytoplasmic accumulation of β-catenin which subsequently translocates into the nucleus to influence gene transcription "

So LRP5 is another GSK-3B inhibitor.  Bone vibration also inhibits GSK-3Beta.

"The initial motivation leading to the identification of the LRP5–5-HT–osteoblast pathway was to better understand the skeletal role of LRP5. Bones from mice with a loss-of-function mutation in Lrp5 (Lrp5^−/−) exhibited a specific molecular signature of decreased expression of genes associated with cell proliferation, but not osteoblast differentiation, bone matrix deposition, or osteoclast differentiation "

"Lrp5 gain-of-function mutations exhibited significantly increased femoral bone mass, compared to knock-in mice not harboring the Col2.3-Cre transgene."

As for the contradictory statements throughout the study about Serotonin:

"It is possible that 5-HT has both anabolic and catabolic skeletal effects, with the prevailing effect being determined by the extracellular concentration of 5-HT and the respective 5-HT receptor/s activated."

To grow taller it may be better to modulate LRP5 or find other ways to phosphorylate GSK-3Beta.

"The negative skeletal effects of elevated circulating 5-HT also do not explain the negative skeletal effects of SSRIs. Administration of a single-dose of an SSRI (fluoxetine) transiently increases plasma 5-HT levels; however, chronic administration of the same SSRI over 1–2 weeks results in substantial reductions in both whole-blood and plasma 5-HT levels"

So the negative feedback mechanism results in lowered Serotonin levels overall(cycling could help?).

"SSRIs impact the skeleton by directly inhibiting the 5-HTT located on bone cell membranes. This may increase local 5-HT levels, in spite of decreased circulating 5-HT, by reducing its removal from the bone cell microenvironment."

According to Static and cyclic tensile strain induce myxomatous effector proteins and serotonin in canine mitral valves, static and cyclic tensile strain can induce both serotonin and chondrogenic factors.

Serotonin acts as a receptor for LRP5 which is a GSK-3Beta inhibitor. Even though this study was on bone cells, inhibiting GSK-3Beta has been shown to increase height in chondrocytes.  Inhibition of GSK-3Beta results in enhanced hypertrophic differentiation of chondrocytes.  Unfortunately, there are no direct studies on Serotonin and chondrocytes. Serotonin and Fluoxetine do have the potential to increase height but care has to be involved in terms of addressing the negative feedback mechanisms. Alternatively, other ways of phosphorylating GSK-3Beta can be found or methods to modify LRP5 directly.


Inactivation of Lrp5 in Osetocytes Reduces Young's Modulus and Responsiveness to the Mechanical Loading.

"Low-density-lipoprotein receptor-related protein 5 (Lrp5) is a co-receptor in Wnt signaling, which plays a critical role in development and maintenance of bone. Osteoporosis-pseudoglioma syndrome, for instance, arises from loss-of- function mutations in Lrp5, and global deletion of Lrp5 in mice results in significantly lower bone mineral density. Since osteocytes are proposed to act as a mechanosensor in bone, we addressed a question whether a conditional loss-of-function mutation of Lrp5 selective to osteocytes (Dmp1-Cre; Lrp5(f/f)) would alter responses to ulna loading. Loading was applied [AXIALLY to the right ulna for 3 min (360 cycles at 2 Hz) at a peak force of 2.65 N for 3 consecutive days, and the contralateral ulna was used as a non-loaded control. Young's modulus was determined using a midshaft section of the femur. Compared to age-matched littermate controls, mice lacking Lrp5 in osteocytes exhibited smaller skeletal size with reduced bone mineral density and content. Compared to controls, Lrp5 deletion in osteocytes also led to a 4.6-fold reduction in Young's modulus. In response to ulna loading, mineralizing surface, mineral apposition rate, and bone formation rate were diminished in mice lacking Lrp5 in osteocytes by 52%, 85%, and 69%, respectively.  Loss-of-function mutation of Lrp5 in osteocytes causes suppression of mechanoresponsiveness and reduces bone mass and Young's modulus. Lrp5-mediated Wnt signaling significantly contributes to maintenance of mechanical properties and bone mass."

16-week old mice were used.  Bone images were taken 18 days after the first of three consecutive days in which axial loading was performed.

"the strain gauge was mounted on the lateral face of the ulna, approximately 100 m from the neutral bending axis of the bone"

Osteocyte-specific KO of LRP5 had reduced femoral bone and spinal bone length.

"In response to [a] bout of cyclic loads, the ulna of the KO mice showed significantly higher strain (3,762 ± 69.6 ) than the littermate control mice (703 ± 34.6 )"

"Periosteal mineralizing surface was significantly reduced [in response to LSJL ulnar loading] in the KO mice (31.6 ± 3.1% in control, and 15.2 ± 7.1% in KO).  mineral apposition rate was decreased significantly (0.74 ± 0.15 m/day in control, and 0.11 ± 0.12 m/day in KO)"

"Both global and conditional inactivation of Lrp5 show low bone mass and decreased bone strength. The anabolic response to ulna loading was reduced by ~94% (global KO) and ~78% (conditional KO in this study)."