Stature Gain with Meditation?

In the post about biophotons, Jory expressed his doubts about the possibility of biophotons being able to manipulate height.  Jory has grown several millimeters by lateral synovial joint loading which by best estimation works by mechanotransduction through the actin cytoskeleton of various cell types in the bone(chondrocytes, osteoblasts, and stem cells).  Lateral Synovial Joint Loading works by inducing interstitial fluid flow which brushes past the actin fibers on these cell types causing adaptations.  Several exercises induce interstitial fluid flow but not to the degree that LSJL does; these exercises induce a stature gain that is minor enough not to be detectable.   This interstitial fluid flow works to increase height either by triggering new bone deposition of surface osteoblasts or by organic development of chondrocytes.

Biophotons are a similar signaling mechanism as to mechanotransduction but instead of operating by mechanisms of shear strain, interstitial fluid flow, and hydrostatic pressure; biophotons function based on light and electromagnetic fields.  Like mechanical loading, biophotons have the ability to alter genetic expression.  Which means that biophotons have the ability to increase height.  Like mechanical loading, the genetic expression change generated by things like hypnosis, meditation, etc. may not be enough and more powerful biophoton generating mechanisms may need to be used like Pulsed Electric Magnetic Fields.

Meditation, Hypnosis, Positive Thinking, etc. may be effective in altering biophoton emissions but only to a minor degree and more powerful tools may be needed like PEMFs to get a really effective height or stature gain.

Effect of meditation on ultraweak photon emission from hands and forehead.

"Various physiologic and biochemical shifts can follow meditation. Meditation has been implicated in impacting free radical activity. Ultraweak photon emission (UPE, biophoton emission) is a constituent of the metabolic processes in a living system. Spectral analysis showed the characteristics of radical reactions.
 Recording and analysing photon emission in 5 subjects before, during and after meditation.
UPE in 5 subjects who meditated in sitting or supine positions was recorded in a darkroom utilising a photomultiplier designed for manipulation in three directions.
Data indicated that UPE changes after meditation. In 1 subject with high pre-meditation values, UPE decreased during meditation and remained low in the postmeditation phase. In the other subjects, only a slight decrease in photon emission was found, but commonly a decrease was observed in the kurtosis and skewness values of the photon count distribution. A second set of data on photon emission from the hands before and after meditation was collected from 2 subjects. These data were characterised by the Fano factor, F(T), i.e. variance over mean of the number of photoelectrons observed within observation time T. All data were compared to surrogate data sets which were constructed by random shuffling of the data sets. In the pre-meditation period, F(T) increased with observation time, significantly at time windows >6 s. No such effect was found after meditation, when F(T) was in the range of the surrogate data set.
The data support the hypothesis that human photon emission can be influenced by meditation. Data from time series recordings suggest that this non-invasive tool for monitoring radical reactions during meditation is useful to characterise the effect of meditation. Fano factor analysis demonstrated that the time series before meditation do not represent a simple Poisson process. Instead, UPE has characteristics of a fractal process, showing long-range correlations. The effect of meditation waives out this coherence phenomenon, suggesting a weaker and less ordered structure of UPE. In general, meditation seems to influence the complex interactions of oxidative and anti-oxidative reactions which regulate photon emission. The reason for the statistical changes between pre- and post-meditation measurements remains unclear and demands further examination."

Now the reason that the photon emissions went down could be because they were no longer moving and weren't performing as many redox reactions.  Also, this study seems to suggest that the mind has the power to influence the oxidative reactions themselves.

During meditation, people control their breathing and breathe slowly thus making it less likely that they perform redox reactions.  Meditation did not change the frequency or color of the biophoton emission making the ability of meditation to alter genetic expression unlikely. 

Gene expression profiling in practitioners of Sudarshan Kriya[An Indian Form of Meditation]

"The rapid pace of life, eating habits, and environmental pollution have increased stress levels and its related disorders. The complex molecular response to stress is mediated by stress genes and a variety of regulatory pathways. Oxidative stress is internal damage caused by reactive oxygen species[oxidative stress can cause damage to height related genes]. Increasing evidence suggests that chronic psychosocial stress may increase oxidative stress, which in turn may contribute to aging, and etiology of coronary diseases, cancer, arthritis, etc. Psychophysiological concomitants of meditation have been extensively researched, but there are very little data available on biochemical activity leading to relieving stress by causing a relaxation response by Sudarshan Kriya (SK). SK is a breathing technique that involves breathing in three different rhythms. It is preceded by Ujjayi Pranayam (long and deep breaths with constriction at the base of throat) and Bhastrika (fast and forceful breaths through nose along with arm movements)[You'd expect this sort of activity to increase biophoton emissions].
Forty-two SK practitioners and 42 normal healthy controls were recruited for our study. The practitioners had practiced SK for at least 1 year. Selected normal healthy controls did not perform any conventional physical exercise or any formal stress management technique. Whole blood was used for glutathione peroxidase estimation and red blood cell lysate was used for superoxide dismutase activity assay and for glutathione estimation. White blood cells were isolated from fresh blood and assayed for gene expression using reverse transcriptase-polymerase chain reaction. The parameters studied are antioxidant enzymes, genes involved in oxidative stress, DNA damage, cell cycle control, aging, and apoptosis.
A better antioxidant status both at the enzyme activity and RNA level was seen in SK practitioners. This was accompanied by better stress regulation and better immune status due to prolonged life span of lymphocytes by up-regulation of antiapoptotic genes and prosurvival genes in these subjects[These antiapoptotic genes may express themselves in chondrocytes as well].
Our pilot study provides the first evidence suggesting that SK practice may exert effects on immunity, aging, cell death, and stress regulation through transcriptional regulation."

What's interesting is that you'd effect some of these studies to increase the amount of redox reactions like the fast and forceful breaths.  What is uniform between SK and Meditation is that you are learning to control your breathing.  This could possibly lead to you controlling redox reactions which in turn controls biophoton emissions.  These biophoton emissions increase expression of antiapoptotic genes which would express expression in all areas including growth plate chondrocytes.

So the important thing isn't the meditation, it's the breathing.  Developing breathing control whether through exercise, singing, etc. will probably result in a small stature gain during development by improved biophoton emissions increasing expression of antiapoptotic genes and reducing DNA damage from reactive oxidative species.

Regulation of gene expression by yoga, meditation and related practices: a review of recent studies.

Review study that mentions the following studies:

Genomic Profiling of Neutrophil Transcripts in Asian Qigong Practitioners: A Pilot Study in Gene Regulation by Mind–Body Interaction

"Six (6) Asian FLG practitioners and 6 Asian normal healthy controls were recruited for our study. The practitioners have practiced FLG for at least 1 year (range, 1–5 years). The practice includes
daily reading of FLG books and daily practice of exercises lasting 1–2 hours. Selected normal healthy controls did not perform Qigong, yoga, t’ai chi, or any other type of mind–body practice, and had not followed any conventional physical exercise program for at least 1 year. Neutrophils were isolated from fresh blood and assayed for gene expression, using microarrays and RNase protection assay (RPA), as well as for function (phagocytosis) and survival (apoptosis).
The changes in gene expression of FLG practitioners in contrast to normal healthy controls were
characterized by enhanced immunity, downregulation of cellular metabolism, and alteration of apoptotic genes in favor of a rapid resolution of inflammation. The lifespan of normal neutrophils was prolonged, while the inflammatory neutrophils displayed accelerated cell death in FLG practitioners as determined by enzyme-linked immunosorbent assay. Correlating with enhanced immunity reflected by microarray data, neutrophil phagocytosis was significantly increased in Qigong practitioners. Some of the altered genes observed by microarray were confirmed by RPA.
Qigong practice may regulate immunity, metabolic rate, and cell death, possibly at the transcriptional level. Our pilot study provides the first evidence that Qigong practice may exert transcriptional regulation at a genomic level. New approaches are needed to study how genes are regulated by elements associated with human uniqueness, such as consciousness, cognition, and spirituality."

A spot analysis of the genes revealed no overtly height increasing genes with the exception of some apoptosis genes.

 Genomic Counter-Stress Changes Induced by the Relaxation Response

"We assessed whole blood transcriptional profiles in 19 healthy, long-term practitioners of daily RR practice (group M), 19 healthy controls (group N1), and 20 N1 individuals who completed 8 weeks of RR training (group N2)."

Gene expression ontology mostly revealed that the gene alteration was anti-catabolism.  Nuclear messanger RNA splicing genes were also altered.

A full gene list was not available.

Genome-wide expression changes in a higher state of consciousness.

"We assessed the whole genome gene expression analysis of long-term meditators in four separate trials and detected significant differential gene expression in association with higher states of consciousness."

A full gene list is given.

A spot analysis for any of the meditators looking for genes related to height revealed that HIF1A was downregulated.  No other overtly pro-chondrogenic or pro-height genes were revealed.

Blood samples were taken so pro-chondrogenic genes directly in the bone marrow may not have been detected.

Grow Taller with Arginine

Arginine is an amino acid that is often taken by height seekers in an attempt to grow taller.  Arginine is taken to try to increase growth hormone levels and growth hormone may increase height(GH has been to shown to increase height in GH transgenic mice and in tumors in the pituitary gland plus other homeostatic altering places but those contain several confounding variables that may increase the height other than the GH itself such as altered homeostatic mechanisms in GH transgenic mice).  Arginine also increases height via the Nitric Oxide/Guanyl Cyclase/cGMP pathways.  CARM1(co-activator associated arginine methyltranferase 1) is a critical regulator of chondrocyte proliferation by regulating the arginine methylation of Sox9.  CARM1 disrupts the interaction of Sox9 with Beta-Catenin (which could induce hypertrophic chondrocytes to differentiate into bone) so someone with more CARM1 may grow taller.

L-Arginine is also a key ingredient of Peak Height, Height Maximizer. 

Ibuprofen-arginine generates nitric oxide and has enhanced anti-inflammatory effects. 

"Ibuprofen, a chiral non-steroidal anti-inflammatory drug chemically related to fenoprofen and naproxen, has moderate but definite anti-inflammatory, analgesic and antipyretic properties. Bioavailability of ibuprofen is increased by salification with various salts.  Ibuprofen-arginine [is] of biological interest because l-arginine acts as substrate of the nitric oxide (NO) synthesising enzymes. Using epithelial HeLa cells expressing the endothelial NO synthase we show that ibuprofen-arginine releases NO and that this NO protects against the cytotoxic apoptogenic effects of staurosporine. Ibuprofen-arginine is endowed with enhanced anti-inflammatory effects with respect to ibuprofen, as shown by reduced hind paw oedema, neutrophil infiltration and chondrocyte apoptosis in collagen-induced mouse arthritis, a model of chronic inflammation. NO has pleiotropic beneficial effects that may contribute to limit inflammation and anti-inflammatory compounds able to release NO display higher efficacy than the parent drugs in defined clinical settings.  NO generation contributes to the enhanced anti-inflammatory effects of ibuprofen-arginine vs. ibuprofen." 

Ibuprofen-arginine may be a potential way to grow taller by increasing NO expression which increases cGK II expression. Nitric Oxide may have negative effects on growth too but this can be alleviated by TP508. 

Thrombin peptide TP508 prevents nitric oxide mediated apoptosis in chondrocytes in the endochondral developmental pathway. 

"TP508 is a 23-amino acid peptide derived from human prothrombin that increases cartilage matrix production and reduces alkaline phosphatase activity without changing chondrocyte proliferation. TP508 acts by blocking the onset of apoptosis associated with hypertrophy. Rat costochondral resting zone chondrocytes and human auricular chondrocytes were cultured in DMEM containing 50microM ascorbic acid and 10% FBS. Apoptosis was induced by treatment of confluent cultures with chelerythrine, tamoxifen, or inorganic phosphate (Pi) for 24h[chelerythrine, tamoxifen, and inorganic phosphate are bad for growth]. One half of the cultures received TP508 (0, 0.7, or 7microg/ml). Apoptosis was assessed as a function of DNA fragmentation ([3H]-thymidine labeled DNA fragments), TUNEL staining, and cell viability using the MTT assay, as well as by assessing the Bcl-2/Bax mRNA and protein ratios and caspase-3 activity. The universal NO synthase inhibitor l-NMMA was used to assess the effect of NO production on chondrocyte apoptosis and specific NO synthase subspecies were identified using iNOS inhibitor 1400W and nNOS inhibitor vinyl-l-NIO, as well as l-NAME, which inhibits both iNOS and eNOS. Finally, we assessed if TP508 would block NO production induced by the apoptogens. Chelerythrine, tamoxifen and Pi-induced apoptosis and this was reversed by TP508. All apoptogens increased NO production and this was reduced by TP508. TP508 reduced NO levels to the same extent as 1400W but not to the same extent as l-NAME, suggesting that its effects are mediated primarily by iNOS. In addition, TP508 reduced the effect of chelerythrine to the same extent as 1400W and l-NAME, again indicating that it acts via inhibition of an iNOS pathway. TP508 also regulated Bcl-2/Bax[mostly via upregulation of Bax] mRNA in a time and dose-dependent manner. The Bcl-2/Bax mRNA ratio was 0.11 in the absence of TP508 at 1h and 4.95 at 7microg/ml TP508; by 3h the ratio was approximately 1 in both groups. The Bcl-2/Bax protein ratio also increased by 63% at 1h. TP508 did not affect caspase-3 activity. TP508 also caused a dose-dependent increase in protein kinase C (PKC) activity within 9min that was maximal at 270min.  TP508 prevents apoptosis in growth plate chondrocytes via inhibition of iNOS-dependent NO [with] PKC [involvement]." 

NO activity may result in chondrocyte apoptosis but NO also has positive activities as well.  TP508 inhibits the specific pathway that causes growth plate chondrocyte apoptosis.  TP508 inhibits apoptosis in human chondrocytes as well.

L-Arginine should increase Nitric Oxide levels by providing more raw materials to increase Nitric Oxide(Nitric Oxide should be increased first by a mechanism like Viagra or Tribulus Terristris).  Viagra might be best because it doesn't increase NO but rather it's a PDE5 inhibitor so it doesn't cause Chondrocyte Apoptosis.

Growing taller with a supplement that increases NO(or inhibits PDE5), L-arginine, or TP508 should be possible.  If you take something like Viagra which doesn't specifically increases NO but rather targets a by-product on the pathway then you may not need TP508.

Mental Height Increase with Biophotons?

Previously, in an article about growing taller with hypnosis, I reported on biophotons being a possible mechanism for how the brain can influence height growth.  We learned that biophotons are stored in DNA and that thoughts can produce electric and magnetic energy which creates an ordered flux of photons.  Pulsed Electric Magnetic Fields alter the expression of certain genes and that expression may be via biophotons.  Maybe it's possible to generate similar magnetic fields only with the power of thought(PEMFs were found to be able to increase cellular proliferation and differentiation).

Biophotons may be a way to control cell growth in cancer and as a corollary may be able to increase cellular proliferation in certain areas(like the growth plate).  Certain genes affect cell growth so for instance a certan biophoton frequency may inhibit myostatin thus stimulating cell growth and another biophoton frequency may upregulate myostatin which would inhibit cell growth.

Biophotons(biological photons) are different colors on the electromagnetic spectrum so thinking with certain colors may be a way to mentally induce height growth by thinking certain colors that upregulate certain genes(biophotons travel through neurons). 

Estimation of the number of biophotons involved in the visual perception of a single-object image: biophoton intensity can be considerably higher inside cells than outside.

"The retina transforms external photon signals into electrical signals that are carried to the V1{You can detect the biophoton signatures of other people; if you often see tall people will you pick up their tall biophoton emissions and grow taller?} (striatecortex). V1 retinotopic electrical signals (spike-related electrical signals along classical axonal-dendritic pathways) can be converted into regulated ultraweak bioluminescent photons (biophotons) through redox processes within retinotopic visual neurons that make it possible to create intrinsic biophysical pictures during visual perception and imagery. biophotons are not by-products [of cellular metabolism], other than originating from regulated cellular radical/redox processes. The biophoton intensity can be considerably higher inside cells than outside. The real biophoton intensity in retinotopic neurons may be sufficient for creating intrinsic biophysical picture representation of a single-object image during visual perception."

So the external biophotons of those around you could possibly alter your own biophoton emissions.  Your retina detects external photon signatures and then those photon signatures are transmitted to your cells by neuronal pathways.  Could you also grow taller by looking at certain frequencies of light? 

Biophoton detection and low-intensity light therapy: a potential clinical partnership.

"Low-intensity light therapy (LILT) [can accelerate] ATP production and [mitigate] oxidative stress. Cellular reduction/oxidation (redox) state may play a central role in determining sensitivity to LILT and may help explain variability in patient responsiveness. In LILT, conditions associated with elevated reactive oxygen species (ROS) production, e.g. diabetic hyperglycemia, demonstrate increased sensitivity to LILT.  The production of [biophotons] is associated with cellular redox state and the generation of ROS."

Cellular reduction/redox state is what determines how sensitive a cell is to biophotons.  How sensitive are chondrocytes? 

Low oxygen reduces the modulation to an oxidative phenotype in monolayer-expanded chondrocytes.

"Autologous chondrocyte implantation requires a phase of in vitro cell expansion, achieved by monolayer culture under atmospheric oxygen levels[chondrocytes like low oxygen environments]. Chondrocytes reside under low oxygen conditions in situ and exhibit a glycolytic metabolism. oxidative phosphorylation rises progressively during culture, with concomitant reactive oxygen species production. We determine if the high oxygen environment in vitro provides the transformation stimulus. Articular chondrocytes were cultured in monolayer for up to 14 days under 2%, 5%, or 20% oxygen. Expansion under 2% and 5% oxygen reduced the rate at which the cells developed an oxidative phenotype compared to 20% oxygen. However, at 40 +/- 4 fmol cell(-1) h(-1) the oxygen consumption by chondrocytes expanded under 2% oxygen for 14 days was still 14 times the value observed for freshly isolated cells. Seventy-five to 78% of the increased oxygen consumption was accounted for by oxidative phosphorylation (oligomycin sensitive). Expansion under low oxygen also reduced cellular proliferation and 8-hydroxyguanosine release, a marker of oxidative DNA damage. These parameters remained elevated compared to freshly isolated cells. Expansion under physiological oxygen levels reduces, but does not abolish, the induction of an oxidative energy metabolism. Simply transferring chondrocytes to low oxygen is not sufficient to either maintain or re-establish a normal energy metabolism. A hydrophobic polystyrene culture surface which promotes rounded cell morphology had no effect on the development of an oxidative metabolism. The shift towards an oxidative energy metabolism is often accompanied by morphological changes."

Low Intensity Light Therapy(which most likely operates by biophotons) has been shown to reduce oxidative stress.  Maybe the low oxidative levels found in chondrocytes is due to the high oxygen consumption of them(the tendency to observe chondrocytes under hypoxia is due to chondrocytes liking to consume oxygen not by them liking to grow under hypoxic conditions).  Low oxygen does reduce the likelihood to acquire an oxidative phenotype and maybe an oxidative phenotype reduces height growth(but low oxygen does induce DNA damage). 

"The increased oxygen consumption of expanded cell populations cannot be accounted for by the utilization of existing mitochondrial reserve function. Rather, the maximal oxidative capacity of these cells is increased[Maybe chondrocytes can adapt to non-hypoxic[above 5% oxygen] environments]. Mitochondrial biogenesis [occurs] during monolayer culture of primary chondrocytes. Together, these data suggest a fundamental adaptation of the chondrocytes towards an oxidative metabolic phenotype, with significant implications for the functionality and longevity of the resulting cell population. Mitochondria [is] a key source of ROS generation. Excessive ROS generation [augments] the formation of altered bases such as 8-hydroxyguanosine in the cellular DNA and promote premature proliferative senescence in a wide range of cells{why reactive oxidative species can reduce height growth and therefore why anti-oxidants can enhance height growth}"


"By culturing chondrocytes using systems which inhibit cell adhesion and maintain a rounded cell morphology, the loss of native synthetic phenotype is inhibited[chondrocytes stay chondrocytes]"<-inhibiting chondrogenic cell adhesion[you need mesenchymal adhesion though to initially form the growth plate] may help height growth by preventing chondrocytes from acquiring a fibroblastic phenotype.

"culture under low oxygen conditions is reported to influence both the maintenance of the native chondrocytic phenotype and its re-expression following monolayer culture"<-lower than 5% oxygen is important but vascularity is needed too at some stages

Transforming growth factor Beta1 induction of tissue inhibitor of metalloproteinases 3 in articular chondrocytes is mediated by reactive oxygen species.

"Transforming growth factor beta1 (TGF-beta1) stimulates cartilage extracellular matrix synthesis but, in excess, evokes synovial inflammation, hyperplasia, and osteophyte formation in arthritic joints. TGF-beta1 induces tissue inhibitor of metalloproteinases 3 (TIMP-3), an inhibitor of cartilage-damaging matrix metalloproteianases and aggrecanases. In primary human and bovine chondrocytes, ROS scavenger and antioxidant N-acetylcysteine (NAC){Even though NAC is an anti-oxidant you don't want to take it as it inhibits NADPH oxidase which is needed for chondrogenic differentiation} inhibited TGF-beta1-induced TIMP-3 mRNA and protein increases. Ebselen and ascorbate also reduced this induction. TGF-beta1 time-dependently induced ROS production that was suppressed by NAC. Hydrogen peroxide, a ROS, induced TIMP-3 RNA. The TIMP-3 increase induced by TGF-beta1 was partly Smad2-dependent. TGF-beta1-stimulated Smad2 phosphorylation was inhibited by NAC{Another reason why NAC is bad for height growth Smad2/3 phosphorylation help height growth}. Reduced glutathione and L-cysteine also blocked Smad2 and TIMP-3 induction by TGF-beta1, whereas a nonthiol, N-acetylalanine, did not. Smad2 was not activated by H2O2. Smad2 phosphorylation was independent, and TIMP-3 expression was dependent, on new protein synthesis. TGF-beta-stimulated ERK and JNK phosphorylation was also inhibited by NAC. However, inhibitory actions of NAC were not mediated by ERK activation. ROS mediate TGF-beta1-induced TIMP-3 gene expression. Blocking TGF-beta1-induced gene expression by modulating cellular redox status with thiols can be potentially beneficial for treating arthritic and other disorders caused by excessive TGF-beta1."

MMP-3 plays a role in the formation of cartilage canals.  Too much vascularity like by FGF or VEGF has been shown to reduce final height growth.  Appropriate levels of TIMP-3 may prevent too much vascularity which has the ability to reduce height gain.  Reactive Oxygen Species which are mediated by biophotons mediate TIMP-3 which may affect height growth.  The problem is that reactive oxygen species are needed so you don't want to shut off their production entirely by a mechanism such as NAC.  You just want to get rid of the excess.

Biophotons are neurological signals that can be induced by mental thought or by lights.  These signals may affect redox reactions which may alter gene expression which could alter height growth.  Maybe mental height increase is possible after all.

Growth Cartilage injury gene expression versus LSJL gene expression

Microarray expression analysis of genes and pathways involved in growth plate cartilage injury responses and bony repair.

"In a rat growth plate injury model, tissue from the injury site was collected across the time-course of bone bridge formation. Four major functional groupings of differentially expressed genes with known roles in skeletal development were identified across the time-course of bone bridge formation, including Wnt signalling (SFRP1, SFRP4, β-catenin, Csnk2a1, Tcf7, Lef1, Fzd1, Fzd2{up}, Wisp1 and Cpz), BMP signalling (BMP-2{up in LSJL}, BMP-6, BMP-7, Chrd, Chrdl2 and Id1), osteoblast differentiation (BMP-2, BMP-6, Chrd, Hgn, Spp1, Axin2, β-catenin, Bglap2) and skeletal development (Chrd, Mmp9, BMP-1, BMP-6, Spp1{up in LSJL as Opn}, Fgfr1{up} and Traf6)."

"An undesirable outcome to growth plate fracture (particularly a Salter–Harris type III or IV fracture) is the bony repair of the injured cartilage at the fractured area."

"studies using a drill-hole injury model in the tibial growth plate of rats have identified sequential inflammatory (days 1–4), fibrogenic (days 3–10), osteogenic (days 7–14) and bone maturation responses (days 14–60) involved in the bony repair of the injured growth plate"

"Twenty-four, 6-week-old male Sprague–Dawley rats underwent experimental drill-hole growth plate injury in the proximal tibia of both hind legs. Under anaesthesia, the growth plate was made accessible after creating a cortical window in the metaphysis using a 2-mm wide dental bur. A central disruption of the growth plate was then made after the bur was passed, via the cortical window, through the entire width of the growth plate, into the epiphyseal region. Six rats were sacrificed by a carbon dioxide overdose 1, 4, 8 and 14 days post injury. "

"genes encoding collagens during the early stages of bone bridge formation (day 4 versus day 8) which were down-regulated [included] Col9a1{up} (− 43.67), Col2a1{up} (− 18.28) and Col9a3{up} (− 8.00)"

"Genes up-regulated at this time with known roles in bone formation included Ltbp2 (9.91), Bglap2 (8.00) and Wisp2{up} (7.51)"

"Clqtnf3 {up} was down-regulated (− 54.05) at day 8 versus day 14"

Genes downregulated on day 4 versus day 8 also downregulated in LSJL:

Ccdc49

Genes upregulated on day 4 versus day 14 also upregulated in LSJL:

THBS4

Cma1

Genes downregulated on day 4 versus day 14 also downregulated in LSJL:

c1qtnf3{up}

Slc38a4{up}

Il1rl1{up}

Bcl11b

Sct

Genes downregulated on day 8 versus day 14 also downregulated in LSJL:

C1qtnf3{up}

Slc38a4{up}

Masp1{up}

Aebp1{up}

"(E) Before dissection of the injury site, with red line denoting the pre-determined area to be excised and collected. (F) Injury site after laser capture microdissection. Dashed lines (green) have traced the uninjured adjacent growth plate."

Injury responses and repair mechanisms of the injured growth plate.

"Following an injury, inflammatory, fibrogenic, osteogenic and bone-bridge maturation repair phases have been observed on days 1-3, 3-7, 7-14 and 10 onwards, respectively. Important roles of several growth factors and cytokines (such as PDGF-BB, FGF-2{up}, TNF-alpha? and IL-1beta) have been highlighted, regulating different phases of growth plate injury repair. While intramembranous ossification is the major mechanism responsible for the bony repair, endochondral ossification, to a lesser extent, also plays a role."

"the gene expression of rat neutrophil chemokine CINC-1 (similar to human interleukin-8) significantly increased during the peak of the inflammatory phase (day 1)"

"p38 mitogen activated protein kinase (MAPK) increased in activation at the injured growth plate"<-lsjl activates p38 MAPK.

"TNF-alpha [has] a strong chemotaxis role for mesenchymal stem cell migration during wound repair"<-TNFa gets stem cells to the wound site.

"neutrophil[involved in inflammatory stage] depletion decreased the expression of chondrogenesis-related genes such as Sox-9 and collagen -2"

"neutrophils [helps initiate] the growth plate injury response and may enhance chondrogenic differentiation"

"Injury-induced inflammatory response[by Cox2 and iNos] in general at the growth plate injury site is necessary for enhancing the chondrogenic differentiation of mesenchymal cells. "<-LSJL upregulates Cox2 as Ptgs2 by 7.63 fold.

"During the influx of fibrogenic cells in both injured growth plate and bone, mRNA levels of growth factors FGF-2 and PDGF-BB [were] significantly upregulated, indicating the possible involvement of both growth factors in regulating this mesenchymal reaction phase in both bone or growth plate injury repair "<-LSJL upregulates FGF2.

"During bone fracture repair, PDGFs have been found to be essential for triggering the initial events leading to the migration and proliferation of fibroblasts and osteoblasts "

Microarray analysis of irradiated growth plate zones following laser microdissection shows later importance of differentially expressed genes during radiorecovery.

"5-week-old male Sprague-Dawley rats underwent fractionated irradiation to the right tibiae over 5 days totaling 17.5 Gy and were then killed at 7, 11, and 16 days following the first radiotherapy fraction. The growth plates were collected from the proximal tibiae bilaterally and subsequently underwent laser microdissection to separate reserve, perichondral, proliferative, and hypertrophic zones.

The reserve zone showed the greatest number of differentially expressed genes and enriched pathways: 259 and 134, respectively. Differentially expressed genes included: Timp3, Gpx1, Gas6, Notch2, VEGF, and HIF-1. Enriched pathways included the developmental processes of regeneration, antiapoptosis, developmental growth, tissue regeneration, mesenchymal cell proliferation, negative regulation of immune response, and determination of symmetry. The reserve zone late upregulation of genes was validated using real-time PCR for Mgp, Gas6, and Eef1a1{down}.

A significant difference in late upregulated genes between growth plate zones exists. The reserve zone shows the greatest change, containing a 10-fold increase in the total number of genes differentially expressed between days 7 and 16. These findings suggest that reserve zone chondrocytes may play a later role in growth plate recovery response following irradiation."

Genes upregulated at day 16(when radiation recovery occurred) that are also upregulated by LSJL:

Col3a1

MMP2

Col12a1

Agc1

Lox

Col10a1

Bgn

Col6a3

Eef1a1{down}

Kras{down}

Col16a1

23.7% of genes upregulated on day 16 were also expressed by LSJL.  The genes above are associated with the reserve zone.

Proliferative Zone:

Bsp

Col5a2

Col1a1

Lum

Kifc1{down}

ADAMTS1

56%.

Hypertrophic Zone:

Rsad2{down}

Ccng2{down}

"blocking KDR{up by LSJL} resulted in the inhibition of VEGF, supporting the concept that the migration and invasion of endothelial cells are regulated by VEGF produced by chondrocytes via a paracrine angiogenic loop"

"Ednra has been shown to bind to endothelin 1{up in LSJL} and induce an increase in intracellular calcium"

Articular cartilage injury:

FGF2 drives changes in gene expression following cartilage injury in vitro and in vivo.

"One important injury-activated pathway involves the release of pericellular fibroblast growth factor-2 (FGF2) from the articular cartilage. Using a novel model of murine cartilage injury, and joints from surgically destabilized mice we examined the extent to which FGF2 contributes to the cellular gene response to injury. Femoral epiphyses from 5 week old wild type mice were avulsed into serum-free medium. Explant lysates were western blotted for phospho-ERK, phospho-p38 and phospho-JNK or were fixed for immunohistochemistry for nuclear translocation of p65 (indicative of NFκB activation). RNA was extracted from injured explants, rested explants stimulated with recombinant FGF2 or FGF18, or whole joints of either wild type or Fgf2-/- mice. RT-PCR was performed for a number of inflammatory response genes previously identified from a microarray analysis. Murine cartilage avulsion injury resulted in the rapid activation of the three mitogen activated kinase pathways as well as NFκB. Almost all genes identified in murine joints following surgical destabilization were also regulated in cartilage explants upon injury. Many of these genes, including activin A, TNF-stimulated gene 6, MMP19, tissue inhibitor of metalloproteinase 1 and podoplanin were significantly FGF2 dependent following injury to cartilage in vitro and to joint tissues in vivo.  FGF2-dependent gene expression occurs in vitro and in vivo in response to cartilage/joint injury."

"inhibin βA (the subunit of activin A), TNF-stimulated gene 6 (Tsg6) and arginase 1, [are inducted] over the first 4h following cartilage injury."

Height Increase Progress Update: Alternating Limbs may be Key

Some people have reported unilateral growth including Richo and his bone length growth.  I've been focusing more on my left leg since it's the one that I've been using for measurements(I'm right handed so I wanted to bring up my weaker side) and I wanted to see too if I had been getting unilateral growth.

My left ankle is bigger and my left leg is a little bit longer.  The overall bone quality is much higher for my left leg.  My routine involves 30 seconds with a 75 lbs dumbell and 30 seconds of tapping with a 10 lbs dumbell(Read more about this Height Increase Method).  With this each day I would alternate starting with my left side and starting with my right side(i.e. one day I would start by loading my left ankle with a 75lbs dumbell and the next day I would start by loading my right ankle with a 75lbs dumbell).  With the table clamp I would always load my left limb first.  Now I am going to be alternating days of starting with the right and left side to see if it makes a difference.

Why would it make a difference?  In response to loading, adaptions to the actin cytoskeleton occur.  These adaptations to the actin cytoskeleton lower the mechanical sensitivity to load.  These adaptations occur rapidly.  By loading one limb first, the actin cytoskeleton may adapt lowering the effectiveness of the loading regime on the other limb.

Here's an old email I got from Hiroki Yokota about local versus systemic effects:

"Your point about systemic vs. local effects is a good question. We used two types of controls – one control is a separate animal without any loading, and the other control is a contralateral control in the same animal (e.g., a right leg is loaded, and a left leg is non-loaded as a contralateral control). These studies indicate that joint loading has a strong local effect compared to a contralateral control. However, a contralateral control seems to receive some benefit of loading compared to the other control animal that does not receive any loading. Thus, a working hypothesis is that joint loading provides significant local effects on the loaded limb (e.g., loaded femur and tibia in response to knee loading), but it can induce systemic effects. The mechanism for systemic effects can be potentially mediated by circulating molecules (as you suggest like growth hormone)."

 So loading on one limb can have an impact on the unloading limb(including possibly remodeling of the actin cytoskeleton).  Muscle too responds via the actin cytoskeleton so we can see if there are similar adaptations in the contralateral limb after doing say bicep curls with one limb.  We will have to investigate this further and could possibly use muscle studies as a basis.


The ramifications:  Each day alternate between limbs(Right limb only, left, etc.) or alternate which limb you load first.

Sprout Taller with Platelet-Derived Growth Factor

In our analysis of F-spondin, we found platelet-derived growth factor was one of the compounds affected by F-spondin.  Inhibiting F-spondin increased height by 30%.  So is inhibiting Platelet-derived growth factor the method by which inihibiting F-spondin increases height?  The most likely mechanism so far is that F-spondin acts by inhibiting FGF(specifically FGF-3).  You need a little bit of FGF but not too much and inhibiting F-spondin may provide a limiting factor on FGF growth.  Still, can augmenting your platelet-derived growth factor levels make you sprout taller?

Regulation of chicken ccn2 gene by interaction between RNA cis-element and putative trans-factor during differentiation of chondrocytes.

"CCN2/CTGF[Connective Tissue Growth Factor] is a multifunctional growth factor. CCN2 plays important roles in both growth and differentiation of chondrocytes and that the 3'-untranslated region (3'-UTR) of ccn2 mRNA contains a cis-repressive element of gene expression.  The stability of chicken ccn2 mRNA is regulated in a differentiation stage-dependent manner in chondrocytes. Stimulation by bone morphogenetic protein 2, platelet-derived growth factor, and CCN2 stabilized ccn2 mRNA in proliferating chondrocytes but that it destabilized the mRNA in prehypertrophic-hypertrophic chondrocytes. The minimal repressive cis-element[a DNA regulatory element] of the 3'-UTR of chicken ccn2 mRNA was located within the area between 100 and 150 bases from the polyadenylation tail. The stability of ccn2 mRNA was correlated with the interaction between this cis-element and a putative 40-kDa trans-factor in nuclei and cytoplasm. The binding between them was prominent in proliferating chondrocytes and attenuated in (pre)hypertrophic chondrocytes. Stimulation by the growth factors repressed the binding in proliferating chondrocytes; however, it enhanced it in (pre)hypertrophic chondrocytes. Gene expression of ccn2 mRNA during endochondral ossification is properly regulated, at least in part, by changing the stability of the mRNA, which arises from the interaction between the RNA cis-element and putative trans-factor."

"ccn2-overexpressing transgenic mice presented a dwarfism phenotype and decreased bone density, whereas ccn2-null mice showed skeletal dysmorphisms as a result of impaired chondrocyte proliferation "

So platelet-derived growth factor(and BMP-2) has the potential to decrease height by destablizing DNA in hypertrophic chondrocytes but it's also needed to stimulate chondrocyte proliferation.  So you need an optimal amount of platelet-derived growth factor to achieve your maximum height.

Impaired vascular invasion of Cbfa1-deficient cartilage engrafted in the spleen.

"Cbfal-deficient (Cbfa1-/-)[Cbfa1 is part of the RUNX2 family] mice displayed a complete absence of osteoblast and osteoclast maturation as well as severely inhibited chondrocyte maturation in most parts of the skeleton. Although chondrocyte maturation and mineralization were observed in restricted areas of Cbfa1-/- mouse skeleton, vascular invasion of calcified cartilage was never noted. To investigate the possibility of chondrocyte maturation and vascular invasion in Cbfal-/- cartilage and the role of the hematopoietic system in the process of vascular invasion, we transplanted embryonic day 18.5 (E18.5) Cbfa1-/- femurs, which are composed of immature chondrocytes, into spleens of normal mice. One week later, the transplanted femurs contained terminally differentiated chondrocytes expressing osteopontin, bone sialoprotein (BSP), and matrix metalloproteinase (MMP) 13. In the diaphyses of the transplants, the cartilage matrix was mineralized and the cartilage was invaded by vascular vessels and osteoclasts. However, chondrocyte maturation and vascular invasion were severely retarded in comparison with transplants of E14.5 wild-type femurs, in which the cartilage was rapidly replaced by bone, and neither mature osteoblasts nor bone formation were observed. In primary culture of Cbfa1-/- chondrocytes, transforming growth factor (TGF) beta1, platelet-derived growth factor (PDGF), interleukin (IL)-1beta, and thyroid hormone (T3) induced osteopontin and MMP-13 expression.  The hematopoietic system are able to support vascular invasion of cartilage independent of Cbfal but are less effective without it, suggesting that Cbfal functions in cooperation with factors from bone marrow in the process of growth plate vascularization."

Platelet-derived growth factor enhances MMP-13 expression and growth plate vascularization thus potentially helping form cartilage canals.

Transforming growth factor-beta 1: induction of bone morphogenetic protein genes expression during endochondral bone formation in the baboon, and synergistic interaction with osteogenic protein-1 (BMP-7).

"TGF-betas do not initiate bone formation when implanted in heterotopic (extraskeletal) sites of rodents. Platelet-derived porcine TGF-beta 1 (pTGF-beta 1) induces endochondral bone in heterotopic sites of the baboon (Papio ursinus) at doses of 5 microgram per 100 mg of guanidinium-inactivated collagenous bone matrix as carrier, with an inductive efficiency comparable to 5 and 25 micrograms of recombinant osteogenic protein-1 (hOP-1, BMP-7), a well characterized inducer of bone formation. pTGF-beta 1 and hOP-1 interact synergistically to induce large ossicles in the rectus abdominis of the primate as evaluated by key parameters of bone formation on day 14 and 30. Tissue generated on day 30 by 5 microgram pTGF-beta 1 or 25 micrograms hOP-1 induced comparable expression levels of OP-1, BMP-3 and type IV collagen mRNA transcripts, whereas TGF-beta 1 and type II collagen expression was 2 to 3 fold higher in pTGF-beta 1-treated implants. In ossicles generated by 25 micrograms hOP-1 in combination with relatively low doses of pTGF-beta 1 (0.5, 1.5 and 5 micrograms), type II collagen expression increased in a pTGF-beta 1 dose-dependent manner, whilst type IV collagen was synergistically upregulated with a 3 to 4 fold increase compared to ossicles generated by a single application of 5 micrograms pTGF-beta 1 or 25 micrograms hOP-1. Morphogen combinations (5 micrograms pTGF-beta 1 with 20 micrograms hOP-1, and 5 and 15 micrograms pTGF-beta 1 with 100 micrograms hOP-1 per g of collagenous matrix as carrier) induced exuberant tissue formation and greater amounts of osteoid than hOP-1 alone when implanted in calvarial defects of the baboon as evaluated. on day 30 and 90, with displacement of the temporalis muscle above the defects. Since a single application of TGF-beta 1 in the primate did not induce bone formation in calvarial defects, whilst it induces endochondral bone differentiation in heterotopic sites, the bone inductive activity of TGF-beta 1 is site and tissue specific. mRNA expression of multiple members of the TGF-beta superfamily suggests complex autocrine and paracrine activities of the ligands and different signalling pathways on responding cells during the cascade of endochondral bone formation in the primate."

"specimens of collagenous matrix combined with 1.5 and 5 pg pTGFB1 showed islands of endochondral bone formation as early as day 14, and almost complete ossicles were generated by 5 pg pTGF-Bl on day 30"

"a. bone induction by 1.5 pg pTGF-BI. b, endochondral bone formation by 125 pg hOP-I. c and d, synergistic bone induction by combinations of 25 pg hOP-1 with 1.5 (c) and 5 pg pTGF-fiI (d). Extensive induction of mineralized bone and osteoid, and areas of chondrogenic tissue when using 5 pg pTGF-p1 (d)."

"Tissue morphogenesis and synergistic activity of hOP-l and pTGF-fi1 implanted singly or in combination in the rectus abdominis of the baboon and harvested on day 30. a and b, bone induction by a single administration of 25 bg hOP-l (a) and 5 pg pTGF-BI (b) delivered by 100 pg of guanidinium-inactivated collagenous matrix. c and d, large ossicles generated by 25 pg hOP-l combined with 1.5 (c) and 5 pg pTGF-Bl (d). Areas of chondrogenesis (arrows in c) at the periphery of the newly generated tissue and corticalization of the ossicles"

Platelet derived Transforming Growth Factor Beta has the potential to grow bone anywhere!  This means you can grow new limbs and longer ones at that.  TGF-Beta should be able to grow new cartilage in existing bone as that is not extraskeletal and I think we can see that by LSJL.

Characteristics and regulation of Pi transport in osteogenic cells for bone metabolism.

"Inorganic phosphate (Pi)[Organic Phosphate can be produced by inorganic phosphate and vice versa] is an essential element in the development of osteogenic cells. The translocation of Pi from the systemic to the skeletal extracellular compartment appears to be an important function of osteoblastic cells. The plasma membrane of osteogenic cells is endowed with a sodium-dependent Pi transport system that is regulated by osteotropic factors such as parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP), insulin-like growth factor-1 (IGF-1), platelet-derived growth factor (PDGF) and fluoride. A similar Pi transport system has been recently identified in matrix vesicles derived from the plasma membrane of osteogenic cells, such as epiphyseal chondrocytes or osteoblastic cells. Matrix vesicles are extracellular structures which are considered to play an important role in endochondral and membranous calcification. Pi transport appears to be the driving force responsible for the accumulation of mineral inside the matrix vesicles and thereby can be considered as a pivotal determinant in the induction of the calcification process. Furthermore, modulation of the activity of the Pi transport at the level of the plasma membrane of osteogenic cells by osteotropic factors is transferred to the matrix vesicles derived from these cells. This notion implies that hormonal and other environmental factors, such as Pi itself and calcium, which have a direct impact on the Pi transport activity of osteogenic cells can also influence the capacity of the matrix vesicles to initiate the mineralization of the bone matrix. The cellular mechanisms involved in the regulation of Pi transport by osteotropic factors have been recently investigated. For the PTH/PTHrP regulatory effect, cAMP appears to be the main mediator and the response does not require the de novo synthesis of proteins. For the effects of IGF-1, PDGF and fluoride, tyrosine phosphorylation processes are involved and responses are dependent upon the de novo synthesis of proteins. The molecules responsible for activation of these signaling pathways are currently under investigation. Such an investigation may improve our understanding of the mechanisms underlying the differentiation processes of osteogenesis such as the calcification of the extracellular matrix."

Remember Calcium Phosphate is pretty indistinguishable from Calcium Carbonate aside from that Phosphate group.  Platelet-Derived Growth Factor affects phosphate transport.  Phosphate transport can be the indection of mineralization of the growth plate by mineralizing hypertrophic chondrocytes.  This doesn't have to be a bad thing as more minerals in hypertrophic chondrocytes could determine their final size.  Remember that IGF-1 increases inorganic phosphate transport too...

So platelet-derived factor does help you sprout taller but possibly only in the minimum amounts and inhibition by inhiting F-spondin may increase height as long as you don't inhibit too much.  This is validated by the dwarfism expressed by ccn2 mice.

Thrombopoietic-mesenchymal interaction that may facilitate both endochondral ossification and platelet maturation via CCN2.

"CCN2 plays a central role in the development and growth of mesenchymal tissue and promotes the regeneration of bone and cartilage in vivo. Abundant CCN2 is contained in platelets[More PDGF, more platelets, more CCN2]. In this study, we initially pursued the possible origin of the CCN2 in platelets. First, we examined if the CCN2 in platelets was produced by megakaryocyte progenitors during differentiation. Unexpectedly, neither megakaryocytic CMK cells nor megakaryocytes that had differentiated from human haemopoietic stem cells in culture showed any detectable CCN2 gene expression or protein production. Together with the fact that no appreciable CCN2 was detected in megakaryocytes in vivo, these results suggest that megakaryocytes themselves do not produce CCN2. Next, we suspected that mesenchymal cells situated around megakaryocytes in the bone marrow were stimulated by the latter to produce CCN2, which was then taken up by platelets. To evaluate this hypothesis, we cultured human chondrocytic HCS-2/8 cells with medium conditioned by differentiating megakaryocyte cultures, and then monitored the production of CCN2 by the cells. As suspected, CCN2 production by HCS-2/8 was significantly enhanced by the conditioned medium. We further confirmed that human platelets were able to absorb/uptake exogenous CCN2 in vitro. These findings indicate that megakaryocytes secrete some unknown soluble factor(s) during differentiation, which factor stimulates the mesenchymal cells to produce CCN2 for uptake by the platelets. During bone growth, such thrombopoietic-mesenchymal interaction may contribute to the hypertrophic chondrocyte-specific accumulation of CCN2 that conducts endochondral ossification."

"growth plate chondrocytes facing the bone marrow are known to produce a vast amount of CCN2 in vivo"<-hypertrophic zone.

How R. Venter grew 1/4" with LSJL

R. Venter started Lateral Synovial Joint Loading on July 6, 2010.  His age was 28 and his height was 5'5".  He weight trains 3 times a week which would help by inhibiting myostatin and increasing his GASP-1 levels.  He started out using a 10lbs hammer and loading the ankles and knees for 5 intervals of 20 seconds a piece.  On July 13, 2010 he reported a growth of 2mm.  Starting Week 2 of August he started using the 10" clamp.  He gained another 1 mm on his right leg as of September 1, 2010 and no further gains in his left leg.  As of October 14th, he reported a total growth of 1/4" in his right leg but no further growth beyond the 2 mm in his left.

So in about 3 months R. Venter grew 1/4" but what's interesting is that his left leg did not grow beyond 2mm.

This study here shows the various load pathways involved in LSJL.  LSJL upregulates more genes in the ECM(extracellular matrix) than anything else.  If you look at the growth plates under LSJL you can see the massive amounts of Matrix secreted(the white stuff).  LSJL also increases levels of hyaluronan synthase which has been used as a carrier for BMP-2(which can encourage chondrogenic differentiation).  LSJL also upregulates levels of MMP-3 which is involved in the vascularization and ossification of chondrocytes but also in the formation of cartilage canals.

R. Venter reported that he's always had issues with his left knee possibly due to the fact that his left knee is not secreting extracellular matrix properly.

LSJL can work by multiple mechanisms.  One of the mechanisms could be bone deposition on the subchondral plate resulting in Venter's 2mm gain in his left tibia.  Another could be differentiation of stem cells and periosteal progenitor cells into chondrocytes as a result of TGF-Beta, hydrostatic pressure, and hyaluronan-mediated BMP-2.  These cells then form cartilage canals(thanks to MMP-3) through the perforating fibers of the periosteum, then these cartilage canals turn into new growth plates.  LSJL also inhibits MMP-1 which degrades the extracellular matrix.

The seeming peak that occurs to LSJL could be due to myostatin(inhibited by exercise and other things), telomere length, and DNA Methylation.  Also, cartilage likes hypoxic environments and most adult growth plates are already vascularized.  Exercise increases hypoxia inducible factor-1 which would help cartilage grow.  Also, bone is never completely vascularized allowing some cartilage to grow in the un-vascularized areas.

The diminishing returns of LSJL can also be due to an increase in bone vascularity inhibiting the formation of new cartilage canals.

Loose chondrocytes

Do loose and disorganized chondrocytes have potential to become growth plates?

Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential.

"Loose fragments in osteochondritis dissecans (OCD) of the knee require internal fixation. On the other hand, loose fragments derived from spontaneous osteonecrosis of the knee (SONK) are usually removed. We investigated proliferative activity and redifferentiation potential of normal cartilage-derived and loose fragment-derived chondrocytes.
Cells were prepared from normal articular cartilages and loose fragment cartilages derived from knee OCD and SONK. Cellular proliferation was compared. Redifferentiation ability of pellet-cultured chondrocytes was assessed by real-time PCR analyses. Mesenchymal differentiation potential was investigated by histological analyses. Positive ratio of a stem cell marker CD166 was evaluated in each cartilaginous tissue.
Normal and OCD chondrocytes showed a higher proliferative activity than SONK chondrocytes. Chondrogenic pellets derived from normal and OCD chondrocytes produced a larger amount of safranin O-stained proteoglycans compared with SONK-derived pellets. Expression of chondrogenic marker genes was inferior in SONK pellets. The CD166-positive ratio was higher in normal cartilages and OCD loose fragments than in SONK loose fragments.
The OCD chondrocytes maintained higher proliferative activity and redifferentiation potential compared with SONK chondrocytes. Chondrogenic properties of loose fragment-derived cells and the amount of CD166-positive cells may affect the repair process of osteochondral defects."

Loose fragment cartilage can have high proliferative and redifferentiation capality which bodes well for micro-growth plates.

OCD usually occurs in younger patients whereas SONK occurs in old patients.

"Human chondrocytes isolated from detached OCD fragments maintain similar cell viability to those from healthy normal cartilages. On the other hand, the chondrogenic redifferentiation potential of horse OCD-derived chondrocytes seems to be inferior to that of normal chondrocytes because OCD chondrocytes show lower expression of a chondrogenic transcription factor, Sry-type high-mobility-group box (SOX) 9"

"COL2A1 expression was not detected in monolayer-cultured (dedifferentiated) SONK chondrocytes, whereas type II collagen deposition and COL2A1 expression were observed in tissue samples of SONK loose fragment cartilages"

Hoxa2

Molecular Study of a Hoxa2 Gain-of-Function in Chondrogenesis: A Model of Idiopathic Proportionate Short Stature.

"In a previous study using transgenic mice ectopically expressing Hoxa2 during chondrogenesis, we associated the animal phenotype to human idiopathic proportionate short stature.  This overall size reduction was correlated with a negative influence of Hoxa2 at the first step of endochondral ossification. The persistent expression of Hoxa2 in chondrogenic territories provokes a general down-regulation of the main factors controlling the differentiation cascade, such as Bapx1, Bmp7, Bmpr1a, Ihh, Msx1, Pax9, Sox6, Sox9{up} and Wnt5a. These data confirm the impairment of chondrogenic differentiation by Hoxa2 overexpression. They also show a selective effect of Hoxa2 on endochondral ossification processes since Gdf5 and Gdf10, and Bmp4 or PthrP were up-regulated and unmodified, respectively."



"Sox9 induction relies on Pax1{up} and Pax9 through Bapx1 stimulation"

"Bmp7 null animals present a severe postnatal size reduction"

"Pax9 and Sox6 deficient mice did not feature length reduction, despite the presence of an
impressive phenotype when associated with a loss-of-function of their usual partners Pax1"

Achieve height increase by modifying MMPs?

MMP(or matrix metalloproteinase) are one of the compounds affected by F-spondin and thus could potentially have height decreasing effects.  Zinc stimulates MMP so if MMPs have some height increasing effects then Zinc could help increase height there are also some MMP inhibitors available.  MMP-9 along with VEGF and osteoclasts are involed in chondrocyte ossification.  What are matrix metalloproteinase and how do they affect height? 

Involvement of matrix metalloproteinases in the growth plate response to physiological mechanical load. 

"Enzymes from the matrix metalloproteinase (MMP) family play a crucial role in growth-plate vascularization and ossification via proteolytic cleavage and remodeling of the extracellular matrix. Their regulation in the growth plate is crucial for normal matrix assembly. Endochondral ossification, which takes place at the growth plates, is influenced by mechanical loading. Using an in vivo avian model for mechanical loading, we have found increased blood penetration into the growth plates of loaded chicks. The purpose of this work was to study the involvement of MMP-2, -3, -9, -13, and -16 in the growth plate's response to loading and in the catch-up growth resulting from load release. We found that mechanical loading, as well as release from load, upregulated MMP-2{up in LSJL}, -9, and -13 expressions. MMP-3, associated with cartilage injuries, and its associated protein connective tissue growth factor (CTGF), were downregulated by the load. However, after release from load, MMP-3{up in LSJL} was upregulated and CTGF levels were elevated and caught up with the control. MMP-3 and CTGF were also downregulated after 60 min of mechanical stretching in vitro." 

"In human vein cells, stationary strain significantly increased MMP-2, whereas cyclic strain decreased it. In those same cells, MMP-9 increased in response to stationary strain, but exhibited no response to cyclic strain. Static compression loading on rat caudal vertebrae elevated the expression of MMP-3, but not MMP-13; in bovine synovial cells, MMP-3, but not MMP-1, increased after cyclic tensile strain"

"Connective tissue growth factor (CTGF/CCN2) is regulated by MMP-3 in two ways:
1) it cleaves CTGF protein into two ∼20-kDa fragments that are thought to have distinct functions;
2) it acts intracellularly to enhance CTGF transcription by directly binding to the CTGF promoter. MMP-3 [upregulates] the expression of CTGF in the human chondrocyte cell line HCS-2/8. CTGF promotes proliferation, maturation, and hypertrophy of growth-plate chondrocytes during endochondral ossification. It acts as a central driver of cartilage and bone regeneration. CTGF level is increased in HCS-2/8 cells after exposure to cyclic mechanical force, but is reduced after exposure to cyclic tension force"

"A reduction in growth-plate width, together with an increased number of vessels penetrating the growth plate from the metaphysic, was observed [in response to growth plate stretching]"

"Together with the increase in blood penetration, a quantitative increase in the expression of MMP-2, [-3], -9, and -13 was observed in the [loading] released chicks"<-LSJL is more consistent with release from load.

"CTGF is expressed in the prehypertrophic zone of the growth plate. It regulates chondrocyte proliferation and differentiation by forming a complex with BMP-2. In postfracture regenerating cartilage, its expression is enhanced in the hypertrophic and proliferative chondrocytes, and it is regulated by MMP-3 "

"the process of catch-up growth may resemble that of wound healing."<-Maybe LSJL operates similarly to catch up growth?

Now the increase in MMP-9 could be a result of increased chondrocyte hypertrophy and not the mechanical loading causing premature ossification of the chondrocytes. 

Effects of bisphosphonate on the endochondral bone formation of the mandibular condyle. 

"This study examined the effects of bisphosphonate on the growth of the condylar cartilage. Alendronate (3.5 mg/kg/week) was administered to postnatal day 1 SD rats for 7 and 10 days. The anteroposterior diameter of the developing condyle was unaffected by the alendronate treatment for 7 days. The total thickness of the cartilage layers was also unaffected by the treatment for 7 days. In particular, there was no change in the thickness of the perichondrium and reserve cell layer at the measured condylar regions. The thickness of the proliferating cell layer was reduced significantly, whereas the thickness of hypertrophied cartilage layer was increased. The number of chondroclasts engaged in hypertrophied cartilage resorption was reduced significantly by the alendronate treatment. The level of MMP-9 expression was reduced at both the transcription and translation levels by the alendronate treatment for 7 and 10 days. alendronate (>3.5 mg/kg/week) inhibits the longitudinal growth of the mandibular condyle by inhibiting chondrocyte proliferation and the resorption of hypertrophied cartilage for ossification." 

So MMP-9 may not only have the possibly negative effect of resorping hypertrophic cartilage, it may also promote chondrocyte proliferation. 

Localization of tartrate-resistant acid phosphatase (TRAP), membrane type-1 matrix metalloproteinases (MT1-MMP) and macrophages during early endochondral bone formation. 

"Endochondral bone formation, the process by which most parts of our skeleton evolve, leads to the establishment of the diaphyseal primary (POC) and epiphyseal secondary ossification centre (SOC) in long bones. An essential event for the development of the SOC is the early generation of vascularized cartilage canals that requires the proteolytic cleavage of the cartilaginous matrix[could development of new vascularized cartilage canals lead to new growth plates?]. This in turn will allow the canals to grow into the epiphysis. In the present study we therefore initially investigated which enzymes and types of cells are involved in this process. We have chosen the mouse as an animal model and focused our studies on the distal part of the femur during early stages after birth. The formation of the cartilage canals was promoted by tartrate-resistant acid phosphatase (TRAP) and membrane type-1 matrix metalloproteinases (MT1-MMP). In addition, macrophages and cells containing numerous lysosomes contributed to the establishment of the canals and enabled their further advancement into the epiphysis. As development continued, the SOC was formed, and in mice aged 10 days a distinct layer of type I collagen (= osteoid) was laid down onto the cartilage scaffold. The events leading to the establishment of the SOC were compared with those of the POC. Basically these processes were quite similar, and in both ossification centers, TRAP-positive chondroclasts resorbed the cartilage matrix. However, occasionally co-expression of TRAP and MT1-MMP was noted in a small subpopulation of this cell type. Furthermore, numerous osteoblasts expressed MT1-MMP from the start of endochondral ossification, whereas others did not. In osteocytogenesis, MT1-MMP has been shown to be critical for the establishment of the cytoplasmic processes mediating the communication between osteocytes and bone-lining cells.  Not all osteoblasts transform into osteocytes. MT1-MMP is needed at the very beginning of osteocytogenesis and may determine whether an osteoblast further differentiates into an osteocyte." 

Cartilage canals are formed in existing hyaline cartilage.  To form them you need MT1-MMP(also known as MMP-14) and TRAP.  What exactly is a cartilage canal? 

The role of cartilage canals in endochondral and perichondral bone formation: are there similarities between these two processes? 

"We investigated the development of cartilage canals to clarify their function in the process of bone formation. Cartilage canals are tubes containing vessels that are found in the hyaline cartilage prior to the formation of a secondary ossification centre (SOC). Their exact role is still controversial and it is unclear whether they contribute to endochondral bone formation when an SOC appears. We examined the cartilage canals of the chicken femur in different developmental stages (E20, D2, 5, 7, 8, 10 and 13). To obtain a detailed picture of the cellular and molecular events within and around the canals the femur was investigated by means of three-dimensional reconstruction, light microscopy, electron microscopy, histochemistry and immunohistochemistry [vascular endothelial growth factor (VEGF), type I and II collagen]. An SOC was visible for the first time on the last embryonic day (E20). Cartilage canals were an extension of the vascularized perichondrium[the early form of periosteum] and its mesenchymal stem cell layers into the hyaline cartilage. The canals formed a complex network within the epiphysis and some of them penetrated into the SOC were they ended blind. The growth of the canals into the SOC was promoted by VEGF. As the development progressed the SOC increased in size and adjacent canals were incorporated into it. The canals contained chondroclasts, which opened the lacunae of hypertrophic chondrocytes, and this was followed by invasion of mesenchymal cells into the empty lacunae and formation of an osteoid layer. In older stages this layer mineralized and increased in thickness by addition of further cells. Outside the SOC cartilage canals are surrounded by osteoid, which is formed by the process of perichondral bone formation.  Cartilage canals contribute to both perichondral and endochondral bone formation and osteoblasts have the same origin in both processes." 

So chondrocyte canals actually increased the size of the Secondary Ossification Centers.  VEGF during early development increased height.  Cartilage canals are also formed from periosteum further emphasizing it's importance in height growth.  Therefore, maximizing things like VEGF and MMP-9 during early development may help with height growth by increasing the number of cartilage canals. 

Expression of matrix metalloproteinases during vascularization and ossification of normal and impaired avian growth plate. 

"Enzymes of the matrix metalloproteinase (MMP) family regulate angiogenesis and are involved in the endochondral ossification process. Tibial dyschondroplasia (TD) and rickets are 2 disorders associated with impairments in this process, mainly in the vascularization of the avian growth plate. We induced TD and rickets and studied the expression patterns of 4 members of the MMP family known to be important for endochondral ossification, MMP-2, 3, 9, and 13, in normal and impaired avian growth plates. The expression of MMP-3, 9, and 13 was reduced in the lesions and lined up parallel to the expulsion of blood vessels, which was extended up to the border of the lesion, but did not penetrate into it. Matrix metallopro-teinase-2 was not expressed in the TD lesion but was overexpressed in the rachitic lesion. We also studied the differentiation stage of the chondrocytes populating the lesions and found that the rachitic lesions were populated with proliferative chondrocytes, whereas the TD lesions were filled with chondrocytes that presented both proliferative and hypertrophic markers. MMP-3, 9, and 13 play a role in the vascularization and ossification processes, whereas MMP-2 is related to chondrocyte differentiation and may be involved in cartilage remodeling in the avian growth plate." 

MMP-3, 9, and 13 play a role in the vascularization process and thus in turn the process of forming catilage canals.  But they also play a role in the possibly catabolic process of ossification... 

Akt signaling regulates actin organization via modulation of MMP-2 activity during chondrogenesis of chick wing limb bud mesenchymal cells. 

"Endochondral ossification is initiated by the differentiation of mesenchymal precursor cells to chondrocytes{Note this can occur after fusion}. This process is characterized by a strong interdependence of cell shape and cytoskeletal organization accompanying the onset of chondrogenic gene expression. The activation of matrix metalloproteinase (MMP)-2 [may] be involved in the reorganization of the actin cytoskeleton and that this would require an Akt-dependent signaling pathway in chick wing bud mesenchymal cells. The pharmacological inhibition of Akt signaling resulted in decreased glycosaminoglycan synthesis and reduced the level of active MMP-2, leading to suppressed cortical actin organization which is characteristic of differentiated chondrocytes. The exposure of cells to bafilomycin A1 reversed these chondro-inhibitory effects induced by inhibition of Akt signaling. Akt signaling is involved in the activation of MMP-2 and that this Akt-induced activation of MMP-2 is responsible for reorganization of the actin cytoskeleton into a cortical pattern with parallel rounding of chondrogenic competent cells." 

MMP-2 is crucial for organizing the growth plate by actin based communication.  MMP-2, 3, 9, 13, 14 seem to be important for endochondral ossification.  We already know that Zinc is important.Spring Valley - Zinc 50 mg, 200 Ct. 

Vascular endothelial growth factor (VEGF) induces matrix metalloproteinase expression in immortalized chondrocytes. 

"VEGF (vascular endothelial growth factor), an important angiogenesis factor, appears also to be involved in inflammatory processes. VEGF and its receptors (VEGFR) are expressed on osteoarthritic, but not on normal adult, chondrocytes. The effects of VEGF were studied on immortalized human chondrocytes. Activated matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, tissue inhibitor of metalloproteinases (TIMP)-1, TIMP-2, interleukin (IL)-1beta, IL-6, and tumour necrosis factor-alpha (TNF-alpha) were measured in culture supernatants by enzyme-linked immunosorbent assays, nitric oxide with the Griess reagent, and cell proliferation by [3H]thymidine incorporation. VEGFR-2 mRNA was quantified by real-time reverse transcription-polymerase chain reaction and the protein was identified by immuno-gold electron microscopy. Intracellular signal transduction effects were determined by western blots and electrophoretic mobility shift assays. The chondrocyte cell lines C28/I2, C20/A4, and T/C28a2/a4 expressed functionally active VEGFR-2. VEGF stimulation induced receptor phosphorylation, activation of the mitogen-activated protein kinases ERK 1/2, and long-lasting activation of the transcription factor AP-1 (activator protein-1). VEGF increased secreted MMP-1, MMP-3, and especially MMP-13, which could be effectively reduced by an inhibitor of VEGFR-2 kinase activity. VEGF diminished the expression of TIMP-1 and especially TIMP-2. Under hypoxic conditions, as occur in cartilage, the reduction in TIMP levels was even greater. VEGF induced IL-1beta, IL-6, TNF-alpha, and nitric oxide expression to a small extent and stimulated the proliferation of immortalized chondrocytes. VEGF is an autocrine stimulator of immortalized chondrocytes that mediates mainly destructive processes in osteoarthritis." 

VEGF and MMPs both increase growth by stimulating chondrocyte proliferation even though they increase ossification.  Neither MMP and VEGF inhibition is probably what is causing the 30% height gain in F-spondin inhibition and in fact MMP and VEGF stimulators may increase height growth by increasing chondrocyte proliferation and encouraging the formation of cartilage canals.

Here's something on MMP-3 which is heavily upregulated by LSJL:

Matrix metalloproteinase-3 in articular cartilage is upregulated by joint immobilization and suppressed by passive joint motion.

"Both underloading and overloading of joints can lead to articular cartilage degradation, a process mediated in part by matrix metalloproteinases (MMPs). Here we examine the effects of reduced loading of rat hindlimbs on articular cartilage expression of MMP-3, which not only digests matrix components but also activates other proteolytic enzymes. Hindlimb immobilization resulted in elevated MMP-3 mRNA expression at 6h that was sustained throughout the 21day immobilization period{LSJL does not involve immobilazation, maybe immbolization increases hydrostatic pressure.  Joint movement may help relieve pressure and lack of movement may cause pressure buildup thus LSJL and immobilization may both upregulate MMP-3 by a hydrostatic pressure based mechanism}. MMP-3 upregulation was higher in the medial condyle than the lateral, and was greatest in the superficial cartilage zone, followed by middle and deep zones. These areas also showed decreases in safranin O staining, consistent with reduced cartilage proteoglycan content, as early as 7days after immobilization. One hour of daily moderate mechanical loading, applied as passive joint motion, reduced the MMP-3 and ADAMTS-5 increases that resulted from immobilization, and also prevented changes in safranin O staining. Intra-articular injections of an MMP-3 inhibitor, N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycylhydroxamic acid (NNGH), dampened the catabolic effects of a 7day immobilization period, indicating a likely requirement for MMP-3 in the regulation of proteoglycan levels through ADAMTS-5."

Note though that LSJL involves intermittent loading whereas this, if the immbolization increases hydrostatic pressure hypothesis is correct, is sustained loading thus any damages inferred from immobilization cannot be derived to LSJL due to the fact that with LSJL you get a break from the pressure.

"In cartilage, MMP-3 not only digests many cartilage ECM components, but can also activate the pro-forms of several MMPs and also contribute to the activation of aggrecanase II (ADAMTS-5)"

"we immobilized the right hind limb of rats, which fixed the knee in full flexion"<-this fully flexed state may have increased hydrostatic pressure.

"A significant increase of MMP-3 mRNA expression [occurred] 6 hours after immobilization"<-LSJL is a lot shorter than 6 hours.

MMP-3 though is still the gene that experiences the most substantial upregulation in LSJL.  However, although MMP-3 is catabolic it may be involved in a more remodeling type of adaptation rather than a destructive force.

Age-related changes in the expression of gelatinase and tissue inhibitor of metalloproteinase genes in mandibular condylar, growth plate, and articular cartilage in rats.

"Mandibular condylar cartilage acts as both articular and growth plate cartilage during growth, and then becomes articular cartilage after growth is complete. Cartilaginous extracellular matrix is remodeled continuously via a combination of production, degradation by matrix metalloproteinases (MMPs), and inhibition of MMP activity by tissue inhibitors of metalloproteinases (TIMPs). This study attempted to clarify the age-related changes in the mRNA expression patterns of MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 in mandibular condylar cartilage in comparison to tibial growth plate and articular cartilage using an in situ hybridization method in growing and adult rats. MMP-2 and MMP-9 were expressed in a wide range of condylar cartilage cells during growth, and their expression domains became limited to mature chondrocytes in adults. The patterns of TIMP-1 and TIMP-2 expression were similar to those of MMP-2 and MMP-9 during growth, and were maintained until adulthood. TIMP-3 was localized to hypertrophic chondrocytes throughout the growth stage. TIMP-1{up, heavily as three different isoforms} and TIMP-2 were general inhibitors of MMP-2{up} and MMP-9 in condylar cartilage, while TIMP-3 regulates the collagenolytic degradation of the hypertrophic cartilage matrix."

MMPs are secreted and activated by Zinc ions.

"MMP-1, -8, and -13 are the collagenases that degrade native collagen fibers. MMP-8 and -13 digest type II collagen molecules with relatively high specificity. MMP-2 degrades type IV collagen fibers and denatured collagens, while MMP-9 digests denatured collagens, type IV collagen, and aggrecan"

"the mandibular condylar cartilage consisted of five cell layers with an erosion zone at the ossification front. The five cell layers were: a fibrous layer with fibroblasts embedded in the fibrous connective tissue; a proliferative cell layer with undifferentiated and proliferative polygonal-shaped cells; a transitional cell layer with flattened cells, without lipid drops in the cytosol; a mature cell layer with ovoid shaped chondrocytes, which were differentiated, and had cell polarity; and a hypertrophic cell layer with enlarged cells with disorganized cytosolic structures"

"When the activity of MMP-13 was inhibited indirectly by the application of fibroblast growth factor 2 (FGF2), the thickness of the cartilaginous cell layers increased"

"MMP-2, MMP-9, TIMP-1, and TIMP-2 interact as general proteolytic enzymes and their inhibitors in cartilage metabolism, while MMP-13 and TIMP-3 may specifically contribute to endochondral bone growth."

MMP driven endochondral fracture union proceeds independently of osteoclast activity.

"We examined a number of anti-resorptive treatments to either block osteoclast activity, including the potent bisphosphonates zoledronic acid (ZA) and clodronate (CLOD), which work via differing mechanisms, or antagonize osteoclastogenesis with recombinant OPG (HuOPG-Fc), comparing these directly to an inhibitor of matrix metalloproteinase (MMP) activity (MMI270). Endochondral ossification to union occurred normally in all anti-resorptive groups. In contrast, MMP inhibition greatly impaired endochondral union, significantly delaying cartilage callus removal. MMP inhibition also produced smaller, denser hard calluses. Hard callus remodeling was, as expected, delayed with ZA, CLOD and OPG treatment at 4 and 6 weeks, resulting in larger more mineralized calluses at 6 weeks. As a result of reduced hard callus turnover, bone formation was reduced with anti-resorptive agents at these time points. These results confirm that the achievement of endochondral fracture union occurs independently of osteoclast activity. Alternatively, MMP secretion by invading cells is obligatory to endochondral union."

"the temporal change in MMP-13 expression during fracture repair coincides with the replacement of cartilage by bone.  MMP-9 (gelatinase-9) removes denatured collagen II fragments along with collagens IV, IV and XI."

Slug

Slug is also known as Snail2.

Role of Slug transcription factor in human mesenchymal stem cells.

"We used human MSCs from tibial plateau (TP) trabecular bone, iliac crest (IC) bone marrow and Wharton's jelly (WJ) umbilical cord demonstrating a variability in their mineral matrix deposition, and in the expression levels of TFs including Runx2, Sox9, Sox5, Sox6, STAT1 and Slug, all involved in the control of osteochondroprogenitors differentiation program. Because we reasoned that the basal expression level of some TFs with crucial role in the control of MSC fate may be correlated with osteogenic potential, we considered the possibility to affect the hMSCs behaviour by using gene silencing approach without exposing cells to induction media. Slug-silenced cells changed in morphology, decreased in their migration ability, increased Sox9 and Sox5 and decreased Sox6 and STAT1 expression. On the contrary, the effect of Slug depletion on Runx2 was influenced by cell type. Interestingly, we demonstrated a direct in vivo regulatory action of Slug by chromatin immunoprecipitation, showing a specific recruitment of this TF in the promoter of Runx2 and Sox9 genes."

"Slug-repressed cells had an impaired ability to close the wounded area compared with control and scrambled cells. After 48 hrs, gap closure in Slug-repressed hMSCs was significantly reduced because migration from the border of the wound was very slow"

"STAT1 inhibits chondrocyte proliferation and regulates bone development"

Chondrogenic potential of human mesenchymal stem cells and expression of Slug transcription factor.

"high basal levels of Slug prevent the chondrogenic differentiation of hMSCs, even in the presence of transforming growth factor-β and elevated levels of Sox9."

"In all osteo-differentiated hMSCs, Slug expression significantly increased both at the mRNA and protein level"

"a physical stimulus, such as the pellet 3D culturing technique, may in itself negatively affect expression of Slug. "<-But the 3D cultural technique is not sufficient to reduce Slug expression.

Angptl1 may supress Slug expression and Angptl1 is upregulated by LSJL.

Angiopoietin-like protein 1 suppresses SLUG to inhibit cancer cell motility

"ANGPTL1 suppressed SLUG by inducing expression of miR-630 in an integrin α1β1/FAK/ERK/SP1 pathway–dependent manner."

"ANGPTLs do not bind to the angiopoietin receptor Tie2 or the elated protein Tie1, which indicates that the functional mechanism of ANGPTL proteins may be different from that of angiopoietins"

"ANGPTL1 interacts with integrin α1β1 and represses its downstream signaling"