Monthly Archives: January 2014

Can MSCs form new growth plate?

One of the issues behind neo-growth plate formation in adults is that adult MSCs are different from embryonic stem cells.  So can adult MSCs form new growth plates despite being more heterogeneous than embryonic stem cells and having less replicative potential?  Are the stem cells that become the epiphyseal growth plate special and have effects that cannot be replicated by adult mesenchymal stem cells?

Common Skeletal Growth Retardation Disorders Resulting from Abnormalities within the Mesenchymal Stem Cells Reservoirs in the Epiphyseal Organs Pertaining to the Long Bones

“the key role of the cartilage-bone regions is their responsibility to replenish the physis with committed chondrocytes, during the developmental, maturation and puberty periods.”

“The concentration gradients of substances such as decreased nutrients, decreased oxygen and decreased specific agents, such as NADNicotinamide-adenine-dinucleotide and poly ADP (Adenosine diphophate) ribose, most probably resulting due to the devascularization [affect morphogenic development].”

“The common metabolic features of a cartilaginous tissue are as follows: A tissue poor in cell numbers, a lack of vascularity, lymph and neurits. Also, slow, glycolytic metabolism, low oxygen tension, flagmatish nature and behaviour expressed in cell proliferation, with special capabilities and routes for the synthesis and secretion of extracellular substances. The spatial three-dimensional structure of the tissue is responsible for the tissue function{so maybe another structure could replicate growth plate function?}, rather than the cells themselves. They possess poor capability for wound healing and regeneration; cells present their final differentiation and maturation stages”

“The nutrition of the cartilaginous tissues is mainly by diffusion of small molecular weight
substances as well as limited diffusion of charged molecules. The transport barrier consists of the negative sulfate and carboxylic groups. The collagens and the proteoglycans carry the main mechanical and biomechanical responsibilities, the hydratation-imbibition of the proteoglycans
contribute to the resistance to compressive strains and their distribution, while the collagen fibers are responsible for the tensile strength.”

“cells arrive from a peripheral site of the physis – the so-called La Croix’s ring and Ranvier’s groove, at the interface of the epiphysis – metaphysis –periosteum – perichondrium, as the sanctum of the developmental activities of the precartilaginous stem cells. At these centers of cell populations and along the routes of their migration towards the germ cell zone of the
physis, chemoattractants, receptors and ligands operate in concert, inducing specific recruitment mechanisms”

“Migration assays of cells in vitro by chemoattractants and in vivo migration testing by
an operational approach, installing a flap of tissue as a barrier on the way of cell migrating route, events that lead to a change in their migration direction, almost by 90°, in pendicular to the physis”

“The epiphyses of the long bones behave as organs with specific functions during the developmental processes, responsible for normal skeletal growth, while pathological growth
retardation maladies are expressed as mutations (EXT1&2 and FGFR-3) within the very same cell populations. Among the pathologies the current review will be focused on two of the main
disorders: Osteochondromas, Hereditary Multiple Exostoses (HME) and solitary exostosis, and
Achondroplastic dwarfism; both ailments are initiated and originally expressed in the
mesenchymal stem cell reservoir populations of the epiphyses”

“At the margins of the periphery of the growth plate, at the metaphysis, on the border between
the epiphyses and diaphysis, exist a population of unique cells with the morphology of fibroblasts, present at the early stages of life, populating the structures of the so-called La Croix’s ring and Ranvier’s groove. These cell centers, also named metaphyseal perichondrium, are believed to be the storage sites of the mesenchymal stem cells,
prone to undergo several differentiation steps within the chondrogenic anlage direction. Starting with prechondroblasts and heading towards mature chondrocytes, simultaneously with the migration of the cells from the physeal periphery to the top of the growth plate columns, they replenish the physis with new differentiated – mature chondrocytes.{We have to mimic these events in order to grow taller} These events take place and last all along the developmental stages and ended post puberty, with the closure and the elimination of the physis. The above described events relate to the healthy state of matters. However under
disturbed, pathological circumstances, the cellular differentiative fate and the cellular migration are deviated, leading to extreme clinical manifestations.
a) Formation of bone protuberances perpendicular to the growth plate, the so-called exostoses
as a result of mutations in the EXT tumor suppressing genes coding for glycosyltransferases, responsible for the synthesis of heparan sulfate molecules.  Heparan sulfate molecules in the pericellular sites are known to serve as activators of FGF receptors. These receptors in turn control the metabolic activity of the cells. In states of shortage, or complete lack of heparan sulfate molecules, the direction of cell migration seems to be altered from upward, along the
periphery of the physis, to perpendicular to the growth plate. The final outcome is the
formation of bone protuberances and clinical difficulties including pain, short stature, as
well as a potential to undergo malignant transformation to chondro and osteosarcoma,
since the healthy EXT genes are considered as malignant tumor suppressing genes. Further
details on HME-osteochondromas clinical entities are outlined and differentiated from
the solitary osteochondroma in the next paragraph on neoplastic disorders.
b) Failure of proper function of FGFR3 (fibroblast growth factor receptor 3) (FGF receptor3) with a negative control in response to its ligand FGF-9, expressed as a deviation of the migration of cells. Instead of migrating to the top of the physis columns and elongating the bone height, cells remain in their original vicinity at the physis periphery and cause the typical widening of the epiphyseal head diameter”{how do we induce this migration?}

“In Achondroplasia there is widening of the long bone heads at the metaphyseal regions which is most probably due to the accumulation of cells that fail to migrate due to the mutation in
FGFR3. The responsible cells for both phenomena are the mesenchymal stem cells – the
prechondrocytes in the reservoirs, that fail to migrate along the proper route due to improper
signalling molecules.”

“Improper concentrations of HA, e.g. overproduction induced by Has 2 in the mesoderm, leads to severely malformed –shortened limbs, lacking one or more skeletal elements, with abnormal morphology and inappropriate positioning of the limb elements.
The sustained production of HA perturbs limb development, affecting precartilaginous condensations, chondrogenesis, growth patterning and cartilage differentiation, normal progression and chondrocyte maturation”

“Cell migration tests have been developed by our team in tissue culture plates. Human mesenchymal-precartilagonous cells were seeded to confluence on a half of the plates’ surface,
while an examined agent (chemoattractant) was placed – embedded in a gel of agarose within a
glass ring at the other pole of the plate. Cells from the confluent half migrated towards the other pole pending on the gradient of the treatment. Among the examined agents were TNF γ; IL-LTB-4; c- AMP; MCP-1 (monocytes chemoattractantprotein-1); IL-1; GM-CSF (granulocyte macrophage colony stimulating factor); IL-2; leptin. The last two agents (IL-2 and leptin) accelerated migration of the cultured cells towards the ring containing the chemoattractant.”

“The last two agents (IL-2 and leptin) accelerated migration of the cultured cells towards the ring containing the chemoattractant.”<-Leptin has been established as having a role in endochondral ossification before.  IL-2 will have to be investigated later.

“Leptin affects the chondrogenic axis lineage, especially at the early phase of chondrogenic differentiation, chondrogenic proliferation and differentiation in the epiphyseal growth plate.”

“Leptin is believed to accelerate skeletal growth via the expression of IGF-I receptors.”

“Osteochondromas are usually classified as benign bone tumors, but they are not neoplastic in nature{not growing new tissue}.
They appear to result from aberrant epiphyseal development with displacement of physeal
cartilage through the perichondrial fibrous ring and subsequent growth at right angles to the long axis of the bone.  The lesion is composed of a mature bony stalk that is covered by a cartilaginous cap”

“removal of the ring of La Croix by surgery causes a complete growth arrest of the limbs”<-Would installation of a new ring of La Croix result in a growth restoration?

“Exposure of the periost to TGF-β1 enhances periosteal chondrogenesis”

“Periosteal chondroma is a benign cartilage-forming lesion, located on the surface of the bone, under the periosteum. It is also referred to as juxtacortical chondroma. Priosteal osteosarcoma is a low- to intermediate grade bone-forming sarcoma with predominantly chondroblastic differentiation that develops on the surfaces of long bones in children”

“[These compounds have a migration effect on chondrocytes]: PDGF, IGF-I, TGF-beta, and Hepatocyte growth factor, BMP-2, IL-1 and 2, and calcitonin”

“adhesive molecules and integrins, HA and its cell surface receptors affect cell mobility and migration”

Image of Ring of La-Croix(LC):

Fibroblast growth factor receptor-3 as a marker for precartilaginous stem cells.(Full Study not available)

“The epiphyseal organ contains two kinds of cartilage, articular and growth plate. Both enlarge during the growth phase of life. However, mitosis is not apparent in these tissues. In the current study, a search to trace the reservoirs of stem cells needed for the growth of these cartilages is done. A disorder in which the stem cells responsible for bone growth are mutated is achondroplasia; the mutation resides in the fibroblast growth factor receptor-3. Epiphyses stained with antifibroblast growth factor 3 antibodies reveal clusters of positively stained cells residing in the perichondrial mesenchyme, known as the ring of La Croix. Removal of the ring of La Croix causes a drastic growth arrest in the limbs of rat neonates. Cell cultures derived of the ring of La Croix biopsy specimens show high rates of cell proliferation and cell migration in vitro, in contrast to articular or growth plate derived chondrocytes. These cells stain intensely by antifibroblast growth factor receptor-3 antibodies and antiproliferative cells nuclear antigen, in contrast with articular and epiphyseal chondrocytes. Transfection of cells from the ring La Croix by an adenovirus vector containing the gene encoding for Escherichia coli beta-galactosidase (lacZ), allows tracing of these cells in tissues. Local injections were performed either to the ring of La Croix or to the joint cavity in a guinea pig model. A characteristic distribution was seen after injection. The transfected cells migrated to areas of bone and cartilage formation in the subchondral bone plate and on either side of the growth plate. This labeling and distribution is maintained for as many as 3 months after injection. The cells from the ring of La Croix appear to be responsible for bone growth. Furthermore, perichondrial cells and other precartilaginous cells expressing fibroblast growth factor-3 have been shown to be good cells for implantation to correct defects of articular cartilage.”

CMF608-a novel mechanical strain-induced bone-specific protein expressed in early osteochondroprogenitor cells.

“Microarray gene expression analysis was utilized to identify genes upregulated in primary rat calvaria cultures in response to mechanical force. One of the identified genes designated CMF608 appeared to be novel. The corresponding full-length cDNA was cloned and characterized in more details. It encodes a putative 2597 amino acid protein containing N-terminal signal peptide, six leucine-rich repeats (LRRs), and 12 immunoglobulin-like repeats, 10 of which are clustered within the C-terminus. Expression of CMF608 is bone-specific and the main type of CMF608-positive cells is mesenchymal osteochondroprogenitors with fibroblast-like morphology. These cells reside in the perichondral fibrous ring of La Croix, periosteum, endosteum of normal bone as well as in the activated periosteum and early fibrous callus generated postfracture. Expression of CMF608 is notably absent from the regions of endochondral ossification. Mature bone cell types do not produce CMF608 with the exception of chondrocytes of the tangential layer of the articular cartilage, which are thought to be under constant mechanical loading. Ectopic expression of CMF608 in HEK293T cells shows that the protein is subjected to post-translational processing and its N-terminal approximately 90 kDa polypeptide can be found in the conditioned medium. Ectopic expression of either the full-length cDNA of CMF608 or of its N-terminal region in CMF608-negative ROS17/2.8 rat osteosarcoma cells results in transfected clones displaying increased proliferation rate and the characteristics of less-differentiated osteoblasts compared to the control cells. Our data indicate that CMF608 is a unique marker of early osteochondroprogenitor cells. We propose that it could be functionally involved in maintenance of the osteochondroprogenitor cells pool and its down-regulation precedes terminal differentiation.”

“Mechanical triggering of rat primary calvaria cells was achieved by stretching following irreversible deformation of the cell attachment surface of the culture dish. For the facilitated identification of de novo transcribed genes, nuclear RNA was extracted following 20 min of stretching.”

“genes transactivated following mechanical strain [include] tenascin C, collagen type XII{up in LSJL}, fibronectin, and connective tissue growth factor”

“genes like ADAMTS-1{up}, thrombospondin 1, endothelin-1{up in LSJL} (its activating enzyme was found upregulated), and desmoykin (AHNAK) [are] previously associated with osteoblastic differentiations.”

No other genes from Table 1 were found to be shared between this and LSJL.

“CMF608 expression was significantly increased in response to all the bone formation promoting stimuli applied, the maximal effect being caused by estrogen administration. In contrast, sciatic neurotomy led to down-regulation of CMF608 mRNA levels”

“In situ hybridization analysis revealed peculiar topography and cell specificity of CMF608 expression in the skeletal tissue. The hybridization signal delineated accumulations of fibroblast-like cells in several locations throughout the long bone: perichondral fibrous ring of LaCroix, periosteum, Volkmann’s canals and endosteum”

CMF608 expression

“n situ hybridization analysis of CMF608 expression in normal and fractured rat bones. Hybridization signal is evident as dark dots in the higher magnification brightfield images (C, D, E, F, I), or as white dots in the lower magnification darkfield images (B, H). Expression of CMF608 mRNA in normal rat tibia is shown in panels A–F. Expression of CMF608 in the healing fracture callus 1 week after the fracture is shown in panels G–I. Panels A and B are respectively low power brightfield and corresponding darkfield microphotographs of the metaphysis. Boxed areas, c and d, shown in panel A, are presented at higher magnification in panels C and D, respectively. Hybridization signal for CMF608 concentrates over fibroblast-like cells in the areas of perichondral fibrous ring (C) and metaphyseal periosteum (D). No hybridization signal is detected above progenitor cells in primary and secondary spongiosa. Panel E demonstrates expression of CMF608 in mesenchymal cells within Volkmann’s canal. Panel F shows expression of CMF608 in endosteal fibroblast-like cells in tibial diaphysis. No hybridization signal is detected above osteoblasts. Panels G and H are, respectively, low-power brightfield and corresponding darkfield microphotographs of the area of fracture callus, which is boxed in the inserted image appearing in the right upper corner of panel G. Hybridization signal concentrates over the fibrous part of the callus but not above the cartilaginous or woven bone areas. Panel I is a high power image of the fibrous part of the callus. Hybridization signal is associated with fibroblast-like cells. oc—osteoclast, bm—bone marrow, lc—lining cells, cb—compact bone. Scale bars: A—100 μm; G—200 μm; insert in G—1 mm; C, D, E, F, and I—50 μm.”

” At 3 and 4 weeks postoperation[induction of a fracture], the mature callus was composed mainly of a cancellous bone undergoing transformation into the compact bone with little if any cartilage or woven bone present. Vascularized periosteal tissue was reduced and covered the center of the callus and was entrapped within the newly formed bone. Multiple undifferentiated cells within this tissue continued to show CMF608-specific hybridization signal

“in situ hybridization results suggest that in adult rat bones, activity of the CMF608 gene is confined to a certain subset of skeletal progenitor cells.”

“The presence of a signal peptide, the absence of a transmembrane domain(s), and the presence of structural domains typical for extracellular proteins (leucine-rich and Ig-like repeats) predict that the CMF608 protein product should be secreted out of the cell.”

“[An] important structural feature, present in all the CMF608 orthologous proteins, is conserved integrin recognition RGD motif found, however, outside the 663 amino acid N-terminal fragment. No RGD sequence was identified in adlican. Integrins are signaling receptors that connect the cytoskeleton to the extracellular matrix, and are involved in mechanotransduction. Mechanical stimulation of bone cells by fluid flow induces recruitment of integrins to focal adhesions. Since CMF608 may theoretically serve as a secreted integrin ligand and its expression is responsive to mechanical stimulation, participation of CMF608 in integrin-mediated bone-specific mechanotransduction may be anticipated.”

“Progenitor cells expressing CMF608 have a fibroblast-like morphology and are found at several locations throughout the bone sections. The lack of CMF608 expression in pluripotent C3H10T1/2 mouse mesenchymal progenitor cells allows us to suggest that CMF608 expressing fibroblast-like cells represent already committed skeletal progenitors, whereas their specific localization hints to their involvement in bone modeling by taking part in circumferential growth of the physis (cells within perichondral ring) and bone shaft (endosteal and periosteal cells of diaphysis) as well as the involvement in the reshaping of metaphysis into bone shaft.”<-so less differentiated progenitor cells can be induced to express CMF608 thus becoming growth plate progenitor cells.

In the study Superior Osteogenic Capacity of Human Embryonic Stem Cells Adapted to Matrix-Free Growth Compared to Human Mesenchymal Stem Cells,  CMF608 is expressed in higher levels in embryonic stem cells than human mesenchymal stem cells.

The growth plates does seem to need a specialized stem cell from the Ring of La Croix as no other growth occurs if the ring of la croix is disrupted from delivering cells to the growth plate.  However, there is a specific gene known as CMF608 that can be activated by mechanical loading or fracture.  So it is possible to create these stem cells and it is possible that a LSJL type loading regime can induce the CMF608 gene.

Also the presence of CMF608 in regions other than the ring of LaCroix such as the periosteum, Volkmann’s canal and endosteum suggests that other cells can be used to form new growth plates other than specific zone of LaCroix cells.

Also, a potential height increase method could be to inject CMF608 positive stem cells underneath the periosteum to form newo-growth plates.

If you look at this image of the groove of Ranvier(A) and zone of La Croix(b) you can see that the growth plate chondrocytes seem to spill out of the groove of ranvier.


According to one site, “On the sides of the growth plate (physis) the Ossification groove of Ranvier provides cells for growth in width. The Fibrous Ring of La Croix lies outside the physis and keeps the cells from oozing out under axial loading”

Decreasing BMC for Height Growth?

Demineralized Bone Matrix is used as a scaffold to induce chondrogenesis and mineralized bone is an inhibition of interstitial growth which is how growth plates make you taller.  Demineralized bone matrix binds fibronectin which may be necessary for chondroinduction.  DBM also changes Wnt pathway signaling which may also affect chondroinduction.  In fact, one paper suggests that bone growth on DBM occurs endochondral means which suggests that there is always a cartilage intermediate.  Demineralized bone matrix also induced de novo endochondral ossification when implanted into muscle tissue.

If we could cause the demineralization of bone matrix just in certain spots that may enable the creation of growth plates in those particular areas.

Here’s what demineralized bone looks like:

Divergent Significance of Bone Mineral Density Changes in Aging Depending on Sites and Sex Revealed through Separate Analyses of Bone Mineral Content and Area.

Bone mineral density (aBMD) is equivalent to bone mineral content (BMC) divided by area. We rechecked the significance of aBMD changes in aging by examining BMC and area separately. Subjects were 1167 community-dwelling Japanese men and women, aged 40-79 years. ABMDs of femoral neck and lumbar spine were assessed by DXA twice, at 6-year intervals. The change rates of BMC and area, as well as aBMD, were calculated and described separately by the age stratum and by sex. In the femoral neck region, aBMDs were significantly decreased in all age strata by an increase in area as well as BMC loss in the same pattern in both sexes. In the lumbar spine region, aBMDs decreased until the age of 60 in women, caused by the significant BMC decrease accompanying the small area change. Very differently in men, aBMDs increased after their 50s due to BMC increase, accompanied by an area increase. Separate analyses of BMC and area change revealed that the significance of aBMD changes in aging was very divergent among sites and between sexes. This may explain in part the dissociation of aBMD change and bone strength, suggesting that we should be more cautious when interpreting the meaning of aBMD change.”

So in men Bone Mineral Content increases with age.  A lower bone density means more potential sites of demineralized bone in the bone.  However, since BMC is only measured at certain sites it may be hard to extrapolate to the entire bone.

However, in some cases in aging bone area increases while BMC decreases which would be ideal for creating sites for de novo growth plate formation.

According to Sex-specific relationships between insulin resistance and bone mineral content in Korean adolescents., insulin resistance is linked to decreased bone mineral content. In one study, insulin resistance was linked to increased longitudinal bone growth.  Maybe this is due to insulin resistance causing decreased BMC?

According to Cortical bone is more sensitive to alcohol dose effects than trabecular bone in the rat., alcohol decreases BMC but also decreases other bone parameters.  Ideally, we’d only want a BMC decrease.

In some cases changes in bone architecture may decrease length:

Anti-osteoporotic activity of methanolic extract of an Indian herbal formula NR/CAL/06 in ovariectomized rats.

“Bilateral ovariectomy was performed in female Sprague-Dawley rats under aseptic conditions and the rats were divided into five groups (n=10). Two different doses of methanolic extract of NR/CAL/06 (200 and 400 mg/kg, per oral (p.o.)) were evaluated for anti-osteoporotic activity and raloxifene (5.4 mg/kg, p.o.) was used as a reference standard. Treatment was given for 90 d.

The bilateral ovariectomy in rats resulted in decreased bone strength, bone mineral content and bone weight. The SEM images showed porous, perforated and disintegrated femur bone architecture and decreases in bone length and thickness in OVX rats. These changes were associated with elevated serum levels of calcium, phosphorus and ALP. Increases in body weight and adipose weight and a decrease in uterine weight were also observed and the changes were highly significant when compared with the sham-control group. Treatment with methanolic extract of NR/CAL/06 (200 and 400 mg/kg, p.o.) for 90 d dose-dependently restored the ovariectomy-induced alterations in bone weight, bone mineral content, bone strength, serum calcium, phosphorus and ALP, body weight and adipose weight nearly to normal levels. Furthermore, the SEM images of the femur bones of NR/CAL/06 (200 and 400 mg/kg, p.o.)-treated rats showed reduced pore formation and improved bone compactness compared with the OVX-control group.”

“The NR/CAL/06 is an herbal formulation consists of mixture of three different plant parts namely, Hibiscus rosasinensis (flowers), Cestrum diurnum (leaves){helps Vitamin D synthesis} and Glycyrrhiza glabra (whole plant){possess estrogen-like activity} in the ratio of 1∶1∶1. ”

The rats were 8 to 10 weeks old when the study began.

Femur length was actually slightly higher in OVX-control group than the sham control.  NR/CAL/06 400mg group had longer femur length than both.  The supplemented groups had a much larger variance of femur length than the sham control meaning that how much OVX or the supplements helps with height depends on the individual rat.

Effect of prior treatment with resveratrol on density and structure of rat long bones under tail-suspension.

“effects of resveratrol on bone mineral density (BMD), bone mineral content, and bone structure were examined in the femora and tibiae of tail-suspended and unsuspended rats using X-ray micro-computed tomography (micro-CT). Rats were treated with 400 mg/kg/day of resveratrol for 45 days and half of them were suspended during the last 2 weeks of treatment. Suspension caused a decrease in tibial and femoral BMD and deterioration of trabecular and cortical bone. Bone deterioration during suspension was paralleled by increased bone marrow area, which could be caused by an increase in stromal cells with osteoclastogenic potential or in adipocytes. Resveratrol had a preventive effect against bone loss induced by hindlimb immobilization. In particular, trabecular bone in the proximal tibial metaphysis was totally preserved in rats treated with resveratrol before tail-suspension.”

Resveratrol can delay fusion.

“[Resveratrol] increases epiphysial bone mineral density (BMD) and inhibits the decrease of femur calcium content in ovariectomized rats”<-interesting that this increasing in bone mineral density is in the epiphysis.

Tibia length in mm


Nonsuspended: 41.58 ± 1.14

Suspended: 42.63 ± 0.68


Nonsuspended: 40.90 ± 0.33

Suspended: 40.60 ± 0.57

Resveratrol decreased bone length and eliminated the bone length advantage from the suspended group.  Note that the suspended group had longer bones than the non-suspended group.  Also, remember that growth rate does not necessarily affect total growth.  So this change in growth rate would not necessarily be reflected in adult height.

Cortical BMD was lower in the suspended group thus possibly suggesting an inverse relationship between BMD and height.  The two resveratrol groups had the highest BMD.

Femur length


Nonsuspended: 37.73 ± 0.80

Suspended: 38.38 ± 1.06


Nonsuspended: 38.75 ± 0.56

Suspended: 37.80 ± 0.32

In the femur the resveratrol nonsuspended femur growth was the highest and it was tied for having the highest bone density.

“In the tibial middiaphysis, measurement of marrow area showed an 1.3-fold increase after 2 weeks of tail-suspension compared to control animals. Resveratrol treatment prevented this increase. In the femoral middiaphysis, the increase in marrow area in suspended rats was more pronounced (1.5-fold compared to unsuspended rats) and could not be totally prevented by resveratrol pretreatment (resveratrol-treated suspended rats showed an 1.2-fold increase in marrow area compared to control rats).”

suspended non-suspended bones

“Micro-computed tomography (micro-CT) slices showing femoral distal (a–d) and tibial proximal (e–h) metaphysis. Suspended rats (b, f) present less trabecular bone than control (a, e), resveratrol-treated (c, g), and suspended + resveratrol-treated rats (d, h)”

“There are mainly two sites that regulate bone structure and bone mass by responding to mechanical stress: one is the osteocytes, surrounded by bone tissue and the osteoblasts next to the osteocytes, and the other is bone marrow composed of mesenchymal and hematopoietic cells. Osteoblasts differentiate from mesenchymal cells and osteoclasts from hematopoietic cells”<-our goal with LSJL is to cause chondrogenic differentiation from mesenchymal stem cells.

According to [Relationship between bone mineral content and growth disorders in children with juvenile idiopathic arthritis]., growth inhibition was associated with lower bone mineral density.

Effects of Khaya grandifoliola on red blood cells and bone mineral content in rats.

“The therapeutic efficacy of a crude water extract of Khaya grandifoliola has been established in mice. This study was designed to assess the effect of the extract on the red blood cells and bone for 7 days, 3 weeks and a recovery period of 3 weeks. Daily administration of the extract showed no related adverse effects on the mortality rate, physical appearance or behaviour of the rats. A general pattern of significant increases in the red blood cell (RBC) count, PCV, haemoglobin and plasma iron content was shown by groups administered with extract after 7 and 21 days when compared with control rats. There was a general trend of reduction in the bone minerals determined (Ca, P, Mg and Cu) in the extract administered groups. Significant decreases were observed at the 500 mg/kg concentration. The bone potassium and iron content was significantly increased in rats administered with extract in a dose-dependent manner. There was an observed significant decrease in alkaline phosphatase (ALP) activity in the rats administered with the extract when compared with the control animals. During the recovery period, the haematological indices regressed to values which were still significantly higher than those of the control values. These results indicate that K. grandifoliola has a positive effect on erythropoeisis, but no significant effect on bone mineral contents at therapeutic doses. At extremely high doses and during prolonged administration, it may have an adverse effect on bone minerals.

The red blood cell count of the 500mg/kg is 23% than the control group.  500 mg/kg also reduced phosphorus, calcium, zinc, and magnesium levels but not iron levels nor potassium levels(which were higher than control).  Perhaps this could decrease the tensile strength of the bone making it more susceptible to stretching forces.

Khaya grandifoliola also decreased alkaline phosphate levels.

There haven’t been a lot of studies on khaya grandifoliola but if reducing bone mineral content does help in a height increase routine that involves stretching the bones than compounds derived from khaya grandifoliola will be extremely helpful.

One study suggested that BMC is linked to height and stated that height loss is predictive of osteoarthritis.

The study also stated that “Most of us are the tallest at around 30-35 years old because of the peak bone mass at that time.”

However, BMC is useful when fighting against the forces of gravity in instances such as suspension then BMC content could be a detriment to the stretching of bones.

New GTPS System or Grow Taller Pyramid Secrets Review

New GTPS System or Grow Taller Pyramid Secrets Review

On Jan 28 we received a message by someone who asked us the question “When are you going to publish your article about GTPS?” – It was by a person who calls themselves calisthenics – There was also another person named Sergi who said the same thing. We are guessing that the two people who sent almost the exact same message is probably the same person. The truth is that we have already in the past reviewed the Grow Taller Guru (Review Here) (aka Lance Ward) and the new system he created called the Grow Taller Pyramid Secrets (The 1st Review is Here). What we sort of figured out about this one person who calls himself the GTG aka the Grow Taller Guru is that the main way he has been marketing and advertising himself is through YouTube, which has become a very good platform to create a unique brand and profile. (You can find his profile link here –

His recent series of video posts have been the new program called Grow Taller Pyramid Secrets. It is also  called the VLog or Video Blog or Video Proof Series. He uses the Slogan “GTG (aka Grow Taller Guru) of the UK to the NYC

Here is the listing of measurements he had done to his students over a 8 week course of exercising, stretching, and diet/nutritional coaching.

Week 1 of Harry & James

Grow Taller Pyramid Secrets 1Time of Reading – Roughly half past five pm (5:00 PM). First Reading during the daytime – 12:00 Noon

  • Harry – Shaggy hair, thin, Caucasian. Took off shoes and stood relaxed in front of the stadiometer. Is wearing black & yellow socks. First Reading – Almost exactly 68 inches (5′ 8”) or around 172.6 cms tall.
  • James – Short clean cut hair w/ Glasses, slightly taller, very pale, thin. Also in socks. Measured reading – Exactly 182 cm in height

Week 11 Measurements

Local Time- 4:49 UK Time

  • Harry Measured Height – 174 -174.1 cm – Posture is slightly more rigid, with hands out to the side
  • James Measurement – 183.9 cm – Posture is much more upright this time, puffed up chest

Week 20 Measurements

Time – 12:40 on Sept 8th, (assuming year is 2013) – (also assuming local UK time)

  • Harry Measured Height – 174.6 cm
  • James Measured Height – 185.4-185.5 – posture is more relaxed with chest not puffed out as much.

Week 30 Measurements

Time – Sunday 12:42 on Nov 17th, 2013

  • Harry Measured Height – 175. 8 cm
  • James Measured Height – 185.6 cm

Week 40 Measurement

Grow Taller Pyramid Secrets 2Time is 12:15 for Saturday, 25th of January

  • Harry Measured height – 178.4-178.5 cm
  • James Measured Height – 185.4-185.5 cm

Other things to mention about the videos that were sent.

The guy is selling Ayurvedic Urea which is supposed to be $550/bottle for free. He is giving away a free height chart. In the background is a picture of Sultan Kosen who was at the time still around 8′ 1″.

Our Analysis On The Height Increase Measured

We had previously tried to explain why one of his clients managed to increase his height by upwards of around 3 cm over a 1 year time span. (Read that review Here). The client was named Michael from Singapore. He originally measured at 8 AM at 161.6 cm on Day 1 to 164.3 & 164.4 on Days 260 and Day 300 respectively. In that past post we had only speculated that the height gain from Lance Ward’s exercise program was real, but only for a certain specific range of person. We had guessed that due to Michael’s previous lifestyle of poor eating habits and lack of exercise, his posture was slouching. Once Michael learned to improve his behavior, his posture improved. His overall increase in 3 cm over a 1 year time is completely reasonable, which could be completely explained from vertebral decompression.

There was a guy on YouTube last year called TightSkinFlash who also through daily stretches gained about 4 cms in height. We did a review on him on the post Reviewing A Height Increase Success Story By SkinTightFlash From Youtube, A Lance Ward Supporter

However that reasoning can’t be used for the cases here, because of Harry.

Where James, who is the taller of the students increased by 3.5 cms, Harry, the shorter of the two, really increased his height beyond would should be possible. I think that the primary focus on our analysis should be on Harry, who grew past the range of what could be considered just stretching and disc or spinal decompression.

Grow Taller Pyramid Secrets 3If we analyzed the videos of Harry of when the videos were started and around the end at 40 weeks, if we looked really closely, we should see that there has been a definite increase in height and change. If we took the height of Lance as the standard, assuming that he did not change in height, (also assuming that his footwear did not change in thickness) then we can see that the top of Harry’s head has increased relative to the position of Lance’s head.

Notice how in the first pictures, which we clipped from the 1st video (for Day #1) the top of Harry’s head is around the level of Lance’s mouth/lips level. By the end of the video series, the top of his head has reached the position of Lance’s Nose position. Of course all of the pictures we clipped is from the video series from the GTG Youtube Channel. The pictures are indeed grainy and the angles have changed, but I have personally tried my best to get the two pictures of the two measurements and positions to be as similar as possible. Harry has definitely increased in height from one picture to the other, at least by our judgement.

Grow Taller Pyramid Secrets ReviewWe can only propose some ideas which can never be validated or discredited, at least without real contact with Lance himself.

Here is a list of our explanations.

The most scientific reason is from jut looking at the body shape of Harry. Notice how skinny both of the students are compared to Lance.

Something that is not well known except among people who study bone growth and auxology is that adolescents and kids who still have growth plates grow vertically first, than followed by horizontally. The long bones in a person’s body first gets longer. Once the growth plates disappear, the bones then get thicker, due to periosteal bone cell growth.

If anyone would look at the relative wrist size of say their father, compared to his son, they would notice that almost always the wrist of the father, who might be shorter than his son, is thicker.

Bones get thicker and wider due to bone growth in the radial direction, as opposed to he axial direction when the hyaline cartilage was still separation the primary ossification center and the secondary ossification center. Once a male reaches full bone maturity, they actually start to grow wider, which is what NBA scouts would describe as to very long & skinny NBA prospects body to “fill out”. The difference between a male body in high school and adulthood is very different, even though they might be the same height.

The adult male body increases in muscle density and content. In fact, the muscle density on a male is highest when he is around the 25-35 year range.

From understanding the linear mechanics on how the growth progression of bones go, we can say that due to how skinny the male students are, they have not ‘filled out” yet, suggesting that they have not reached the end of their bone maturity.

Remember also the fact that there is more than just growth plates in the long bones. The irregular vertebrate bones also have a few layers of hyaline cartilage. While the legs and arms may not be getting longer & contributing to the overall height, we should not forget that the torso makes a major contribution to. If we assume that even 5 of the 33 vertebrate bones have a slight layer of cartilage left, they would lead to some extra height.

Based on our previous research to see how do growth plates actually disappear, we’ve found that the growth plates don’t ossify at the same time. While the hyaline cartilage in the distal tibia fuses the earliest, around 16-18, the cartilage in the vertebrate bones might not fuse until 20-22.

That is what I propose are the list of explanations for Harry’s height increase.

This is all of course assuming that the students are not wearing any type of height increasing socks (we did a report on these unique products which give you a full 1 inch of extra height here), have not reduced the stadiometer, or done any other type of trick as a setup beforehand.

As a person who used to perform sleight of hand tricks and practiced to be an illusionist for a few years, who understands how to trick (and/or misdirect) people through stage setups, preparations, and even using shills, I understand the need to be very sceptical and suspicious of people who make very extreme and extravagant claims. I am willing to believe that Lance might have helped these two British lads increase their height, but would their bodies have gone through the same amount of growth, without his help? That would of course require something very experimental unbiased.

I am not sure where we would be able to do some type of double-blind non-biased test using identical twins for real testing. At this point, the only thing we can do is to take his word on faith while be slightly reserved at the same time.

Gloxi Height Enhancer Ingredient Analysis

Michael has provided his thoughts on Gloxi Height Enhancer here.  I also want to note that I could not find any information on promodulin which is mentioned in the video as being a key to grow taller. I did find something on uromodulin which is connection to their calcium thesis.

I wanted to do a detailed analysis on the ingredients I obtained from this page on Gloxi Height Enhancer.

In the analysis on Alfalfa, it was discovered that Alfalfa may result in increased height but only during development.  One study found that some Lectins could encourage chondroinduction, but wheat germ actually encouraged adipoinduction.  Here’s what Michael had to say about Casein Phosphopeptide.  I think the existing research suggests that Casein Phosphopeptide can accelerate and possibly enhance existing endochondral ossification but it doesn’t look like it can induce new growth plates.  Hydrolyzed collagen has been shown to enhanced longitudinal bone growth in some cases.  According to Effects of Chlorella vulgaris on bone marrow progenitor cells of mice infected with Listeria monocytogenes., Chlorella can reduce lead levels of bone marrow cells.  Lead stimulates TGF-Beta signaling of chondrogenesis but inhibits BMP-2 signaling of chondrogenesis.  BMP-2 signaling seems to be preferably over TGF-Beta signaling for enhanced growth.  Thus Chlorella could contribute to longitudinal bone growth benefits by it’s beneficial effects against lead.  Jujube seems to be a typical antioxidant in which I couldn’t find any novel effects.  Anyone know any?

According to Ancordin, the major rhizome protein of madeira-vine, with trypsin inhibitory and stimulatory activities in nitric oxide productions., Madeira-vine could stimulate nitric oxide production.  It has an inhibitory effect on trypsin and trypsin seems to have a beneficial effect on growth.

The remainder of the ingredients are just too common and well studied and sufficient levels are likely already present in your diet.

Based on this evidence, it’s possible that Gloxi Height Enhancer might enhance height growth in growing individuals but will not re-induce height growth in adults unless these ingredients have novel effects that were previously undocumented.

According to [Effects of astragalus and angelica on bone marrow stem cells proliferation and VEGF protein expression in vitro]., Angelica(and astragalus) could increase stem cell proliferation and stem cells would be the foundation for new growth plates.  Also this study: Angelica sinensis polysaccharide nanoparticles as novel non-viral carriers for gene delivery to mesenchymal stem cells., states that an ingredient from Angelica could be used for gene delivery to MSC.  This could be a very powerful effect for inducing new height growth but it’s not possible to be used in this gel.  I couldn’t find much on Chiang Ziong Rhizonia(more common name is Huang Xiong).

The primary ingredient of rhizoma discoreae is diosgenin.  I studied diosgenin but couldn’t find a connection between it and longitudinal bone growth.

The promoting effects of geniposidic acid and aucubin in Eucommia ulmoides Oliver leaves on collagen synthesis.

“collagen synthesis was stimulated by the administration of a hot water extract from the leaves of Eucommia ulmoides OLIVER, Eucommiaceae (Du-Zhong leaves) in false aged model rats. In this paper, we set out to examine the compounds in Du-Zhong leaves that stimulated collagen synthesis in false aged model rats. In experiment 1, a methanol extract of Du-Zhong leaves also stimulated collagen synthesis in aged model rats. An acetone fraction was derived from the methanol extract by silica gel chromatography in experiment 2. The acetone fraction mainly contained iridoides mono-glycosides such as geniposidic acid and aucubin. The administration of geniposidic acid or aucubin stimulated collagen synthesis in aged model rats in experiments 3 and 4 (significance (p<0.05)). The reported pharmacological effects of Du-Zhong leaves, including healing organs and strengthening bone and muscle, are closely related to collagen metabolism. It appears that geniposidic acid and aucubin are the actual compounds in Du-Zhong which caused the effect in our experiments.”

So could possibly have an effect. Since there’s collagen in the skin using this gel could make your skin thicker.

According to Clove (Syzygium aromaticum Linn) extract rich in eugenol and eugenol derivatives shows bone-preserving efficacy., Clove does alter the serum bone levels of things such as Calcium or Phosphate but nothing that could induce new height growth.

Most of the research regarding Ocimum gratissimum deals with it’s beneficial affects against radiation damage.

Finally, there’s Astragalus which has actually been shown to have a connection to longitudinal bone growth.

If you look on the other part of the brochure you can see that they don’t list any novel effects for the ingredients.  I also don’t see any means of how total calcium absorption will increase height.

Biggest Growth Spurt of 24 inches In 3 Months by 12 Year Old Boy from Hodgkin’s Chemotherapy Treatment

Biggest Growth Spurt of 24 inches In 3 Months by 12 Year Old Boy from Hodgkin’s Chemotherapy Treatment

OLYMPUS DIGITAL CAMERARecently I have been reading the Pulitzer Prize Winner book on the history of cancer called The Emperor of All Maladies by Siddhartha Mukherjee. My cousin which has been trained as an oncologist recommend the book to me and I finally managed to buy the version of the book from Amazon and have been spending about hours in the last few days of traveling in Airports and Planes listening to this incredible story on how physicians for the last few millennia have been fighting against this most difficult of diseases.

Some surprising figures I would learn is that 1 out of every 3 people in the USA would be diagnosed with cancer at some point in their life. Every year about 3 million people die from the disease in the world but the CDC reports that it is probably closer to 8 million people die from some version of Cancer each year. The struggle of doctors to fight and try to win this war against cancer is probably going be the defining medical endeavor for our generation, if not this century.

Maybe during the 19th century and the early 20th century, the focus of physicians was on finding ways to fight against infectious diseases like TB, Smallpox, Polio, and Influenza, those type of diseases caused by the bacteria or parasites have been taken care of ever since the advent of Penicillin and various other vaccinations. The defining types of disorders that are affecting this new generation is going to be degenerative diseases as man kind learn how to extend his life expectancy from 65 to maybe to double that. Our great-grandparents didn’t have to worry about Osteoporosis, Arthritis, High Blood Pressure, or Alzeheimers as much as us since their life expectancy was not as long as ours will be. Cancer will be the other major type of disease which we will have to deal with.

Our Main Point

Biggest Growth SpurtIn a certain passage of the audible book Dr. Mukherjee mentioned that after one case of cancer remission for a boy which had his weight reduced greatly to just 50 lbs by cancer, the 12 year old boy managed to grow 2 feet within 3 months after the oncologists managed to find the right type of chemical compound, (usually a type of anti-folate derivative) to stall the multiplication and spread of the cancer cells in his system. In addition, His weight increased by 50%, which does not seem like a lot since a 50% increase from 50 lbs is just 25 lbs to a total of 75 lbs.

(Read the clipping we did of the PDF of the audio book on the right. It was under the chapter called “An Army On The March“)

This type of story is the strangest which I have ever heard of any growth spurts. It is definitely the biggest growth spurt which I have ever heard off. To grow 2 feet in 3 months would mean that the kid would be growing at around over 1/4 inch every single day!!!

Of course we have to remind ourselves that this is a 12 year old boy we are talking about but this rate is still insane to comprehend.

I am not sure how it is possible that a person can grow that fast, even if we take into account catch up growth. I personally would suspect that after his 3 months of intense height increase, the growth plates probably completely ossified, due to over extension of the process in the cartilage.

(It seems that the growth plates, just like almost every other system/unit in the body that has not been infected by cancer has a natural negative feedback loop controller if one variable becomes too high. This is known collectively as Homeostasis. If you over exert the growth plates, they will go through bone maturity at a faster rate. If your growth is stunted, the senescence of the growth plates actually slows down, leading to catch-up growth after the inhibitory effect is removed. It seems that maybe the chondrocytes in the resting zone have some type of internal, intrinsic rate at which they can replicate and enlarge before some receptor or protein in the ECM causes them to stop.)

We have written before a few times about how catch up growth works.

While we were studying the longitudinal diurnal growth patterns of the tibia, we found that the long bones don’t grow in any type of linear pattern, or even predictable variable and/or intermittent pattern. (For Reference, read “Modulation of vertebral and tibial growth by compression loading: diurnal versus full-time loading“). They go through what can only be described as micro-growth spurts which were irregular . We realized from looking at the pattern on how major growth spurts work, there are certain short durations of time where the hypertrophy process of the chondrocytes seem to go crazy.

The Possible Case and the Growth Curve

What we speculate, without going into the deep research to find the exact case, is that based on the growth curve provided by the pamphlets created annually by WHO (World Health Organization) and the NIH (National Institute of Health) that before the treatment, the boy would probably have been around 3 feet tall. Let’s remember what we learned from previous posts before where we both uploaded and analyzed the charts provided by both the CDC and the WHO.

The average (median, NOT mean) American 12 year old boy would be around 150 cm tall with a 1 standard deviation range of 7-8 cm. The information was obtained by typing in the term “Average Height of 12 Year Old Boy” into the Wolfram Alpha Search Engine. (Based on the 2000 CDC Report).

In addition, the average weight (we are again talking about median) would be 40 kg, with an uneven distribution bell curve. It would range as high as 50 kg or as low as 34 kg.

We assume that the boy did not have Hodgkin’s his whole life, but only developed it probably in the last 2-3 years. That meant that his growth would have been stunted only within the last 3-5 years, although there might have been a few cancer cells in his system before that. From what I can personally remember about this particular type of cancer, which is usually called Lymphoma, it is one of the few cancers which is not too fatal and people who get it treated early enough can have a very high life expectancy past diagnosis.

Since the growth rate per year for an American boy between the ages of say 7-12 or 9-12 is about 2-3 inch/year and also correct that for the factor that increased bump in linear growth in long bones for puberty, we would guess that his height around the age of 12 was stunted by around 10-13 inches of height. Obviously the body was trying to catch up.

If this boy did not have his height stunted, the 2 extra feet of height would have pushed him to be almost 7 feet tall. If we we assume that this boy at the age of 12 was about the 12-13 inches shorter than average, which is at 150 cm (or 4′ 11″), than his increase of 2 feet would put his final height to be around 6 feet tall. Of course, that would still seem a little ridiculous. It is hard for us to imagine a 12 and a half year old boy who has been suffering for a few years from Hodgkin’s growing to the height of 6 feet tall. We probably won’t be provided private information about that boy who lived in the early 20th Century, but we guess that if we are to assume that his height was dramatically stunted to less than 4 feet, then his growth to a final adult height (assuming the boy lived to adulthood) would not seem so extreme.

The case of this boy who at the age of 12 with stunted growth growing 24 inches in height in 3 months shows that the potential for increased bone longitudinal growth is much higher than I would have ever predicted. Could we possibly use a mechanism to stunt our growth at the later stages of bone maturity (ages 16-18) for an extra push in bone longitudinal growth to reach a higher adult height from the rebounding effect of catch-up growth?

Reply To Geoffrey Arnold and His Blog The Social Complex

Reply To Geoffrey Arnold and His Blog The Social Complex

We have a tracking program for our website Natural Height Growth and can see most people who quote or link to the website. We found out from the tracking software that Mr. Arnold wrote on a Reddit thread that we on this website were selling a product so we had to be some type of major scam. We are not selling anything of our own, at least right now.

This is what Geoffrey Arnold has said about us, which we found insulting and rude, since he didn’t even give us a chance and look over what we have been doing for the last few years. Maybe the only way to describe this guy is to call him a “Hater”. He feels like he can’t play in the game since the world is discriminating against him and people of his “kind” unfairly so the only way he know how to respond is just to hate and write.

The back story, on what Geoffrey wrote on the specific Reddit Post…

“….This guy actually sent a message to my blog, advertising his site, as if I was going to publish it. I might get around to reading it later, but I’m sure it’s mostly gibberish. Hard to trust anyone who is talking about heightism while selling a product aimed at people who want to be taller.

That would be like getting information about racism from a company that sells skin bleaching cream…”

The clipping is from below…

Geoffrey Arnold

Our intention was to write two different books, one for people who are still growing and another for people who have stopped growing. At this point, we have looked at the science and say that it is possible for us to write one book for people who still have their growth plate cartilage and can possibly stlll grow. We can’t write the 2nd book for people with no cartilage left. We may never be able to write that 2nd book since we have found very little evidence that it would be possible.

There is a store section on the website but if he had look at it carefully, then he would see we don’t have anything of our own to sell. As for everything else, we give them for free. We even had DMCAs from the people at (Company That Will Not Be Named) and GT4I given to stop us from giving their products for free. We are putting our skin into the game and have come close to legal actions against people because we are trying to make the internet and the world a better place. We are trying to change the way the world operations in a fundamental way.

What don’t think that his work is that impressive. He has said that he has written over 1200+ post for the website. We are now at over 1100+. In terms of quantity, we will overwhelm his website soon.

He would have realized that our motives are not just a financial one. We’d love to actually find a solution and make some big profit from it but we keep the site going from google ads. We are a part of the Amazon Affiliate program but that doesn’t earn enough money since people don’t seem to click those links. This website is not as profitable as most people would believe. We are trying to build up a way to monetize the website to let us continue to pay off the monthly operating costs.

I wrote to him before because I thought he cared enough to want to help and maybe try to look for a solution. He sure has a lot of stuff written about heightism. Everything he says is more or less right, but would he would be willing to put in some type of effort to maybe change the system, and help people? If he doesn’t believe there is a solution, that is fine. He doesn’t need to lift up a medical textbook and go through the real research to see what is possible. He has the complete  right to just spend the next 10+ years and keep writing another 3000+ posts about how the world is bias towards short men and short people.

He’ll get a small following of people on Reddit who agree with him and he will probably will get some type of self satisfaction in knowing that he can only be half-fulfilled. He can justify his own existence in his small circle of influence and control. We get it, the way that the ego works to protect itself is to just be right about something. We might not be able to get what we really, REALLY want, but as long as we can sit back, be pessimistic, sceptical, cynical, or pragmatic about how bad the world treats us because of something which we can’t change, as least we can be partially fulfilled and satisfied. Instead of finding self-esteem in knowing that we can change this problem, he finds validation in his ego from knowing that he is “right” since he just talk about how bad the world is. He can be the sceptic who does nothing, but takes no time or effort to look for a way out of the problems that they vent about. 

What he wrote about us I would consider to be unfair since he never made the full effort to read any of our large posts and research extensively. He brushed us off without ever looking at the science. Why? It is the same reason why I have brushed off any type of height increasing pill or potion or herbal concoction sold by websites using the type of design from the 2004-2009 era. It seems to be too crazy and improbably to believe. However, at least I spend the time to look over what the websites have to say. Maybe they do have something of worth to say and something to contribute. He apparently did not take the time to look over what our research is, and dismissed us without ever looking into our content.