Huogo I Formula

Angelica Sinensis is available for sale Solaray - Dong Quai Angelica Sinensis, 550 mg, 100 capsules

Effects of huogu I formula (I) on correlated factors of bone regeneration in chickens with steroid-induced necrosis of femoral head.

The microenvironment of a femoral head recently undergone necrosis may differ from an adult femoral head.

"Forty-eight healthy female Leghorn chickens were randomly divided into control group, model group and Huogu I group, and each group consisted of 16 chickens. At the meantime of model establishment, chickens of the Huogu I group were administrated with decoction, while the model and control group with distilled water by gavage. At the 8th and 16th week after medication, blood samples were obtained for blood lipid detection while both sides of femoral head were harvested for the rest of examinations. .

Compared with the control group, serum levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the model group rose significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the model group dropped prominently. Positive cell counting of Smad7 and RANKL increased dramatically. In contrast with the model group, levels of TC, TG and LDL-C in Huogu I group reduced significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the Huogu I group increased prominently. Indices of Smad7 and RANKL both decreased significantly. Especially at the 8th week, these variations were more significant.

Huogu I formula is effective in promoting repair of necrotic femoral head by regulating the expressions of BMP2, TGFβ(1), Smads and OPG/RANKL of osteoclast in femoral head."

"Huogu Ⅰ Formula (consisting of Poria Cocos, Atractylodes Rhizome, Codonopsis Pilosula, prepared Pinellia tuber, Radix Paeonia Rubra, Ramulus Cinnamoni, Angelica Sinensis, Ligustici Chuanxiong and prepared Rhizome Rehmannia) provided by Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, was decocted into solution which contained crude drug in 1 g/mL. Methyl prednisolone sondium succinate"

The model group refers to the osteonecrosis group, the control had no stimulation, and the Huogo I group is Huogo I +necrosis.

"Eight weeks after steroid administration, the model group showed several histomorphological changes as compared with the control group: thin trabecula, increased number of empty lacuna and adipocyte proliferation and hypertrophia. At the 16th week, collapsed bone trabeculae, necrosis accompanied by hyperplasia in some regions, increased empty lacuna were observed. Moreover, the fat area in marrow cavity was higher than that in the normal group. At the 8th and 16th week after treated by Huogu Ⅰ Formula, trabeculae were arranged in regular and compact shapes, large amount of osteoclast and few osteoclast distributed around them, and the empty lacuna rate and fat area in medullary channel were significantly lower than those in the model group"

"The expressions of BMP2 and TGFβ1 positive staining were observed in chondrocye and osteocyte of femoral head in the control group. At the 8th week, the expressions of BMP2 and TGFβ1 in the model group were significantly lower than that in the control group. Additionally, the difference was even more obvious at the 16th week. Compared with the model group, the expressions of BMP2 and

TGFβ1 in Huogu Ⅰ group were significantly higher. At the 8th week, the increasing trend of the two factors in Huogu Ⅰ group was drastic and even approximate to counterparts in the control group{but not greater than the control group which is what we would hope if Huogo I formula is height increasing}"

"Rhizoma Chuanxiong is effective in improving microcirculation and reducing the blood viscosity; Atractylodes Macrocephala Koidz, Angelica Sinensis, and prepared Pinellia Tuber possess similar effects and may improve the condition of hyperlipidemia and prevent fat from accumulating in marrow cavity."

"Angelica Sinensis possesses effects of up-regulating expressions of BMP2, cbfal, insulinlike growth factors (IGF), and other related genes of osteoblast{many of these genes are pro-chondrogenic also}. We believe that, on one hand, Huogu Ⅰ Formula acts on osteoblast and bone marrow stromal cell to enhance the expression of related genes of osteogenesis. On the other hand, it regulates the lipid metabolism and improves the condition of abnormal blood rheology, leading to the promotion of blood supply of femoral head."<-So Angelica Sinensis is the most promising compound.


Study to establish the role of JAK2 and SMAD1/5/8 pathways in the inhibition of hepcidin by polysaccharides from Angelica sinensis., states that Angelica Sinensis may inhibit the SMAD1/5/8 pathway.  Smad1/5/8 is the BMP pathway signaling.


Role of Bone Morphogenetic Proteins-7 (BMP-7) in the Renal Improvement Effect of DangGui (Angelica sinensis) in Type-1 Diabetic Rats., states that Angelica sinensis may directly activate BMP-7.

Static compression on growth plates

Static compressive loading reduces the mRNA expression of type II and X collagen in rat growth-plate chondrocytes during postnatal growth.

"This in vitro study investigated the effects of loading on the mRNA expression pattern of key molecular components of the growth-plate related to the extracellular matrix (type II and type X collagen) and the PTH-PTHrP feedback loop. Short-term static compressive loading was applied to rat proximal tibial growth-plate explants. Four age groups at specific developmental stages were investigated. The spatial variation in the mRNA expression was compared among loaded explants, their contralateral sham controls, and uncultured growth plates from normal animals. Basic cell metabolism (18S rRNA) was unaffected by load. Results indicated a narrower spatial distribution of mRNA expression of type II collagen throughout the growth plate; similarly, a narrowed distribution of expression of type X collagen was noted in the lower hypertrophic zone of the growth-plate. Mechanical compression influences chondrocytes of the hypertrophic zone to alter their expression of specific genes encoding proteins of the extracellular matrix, while PTH-PTHrP receptor mRNA, a regulatory protein, remained unaffected by loading. The effects of compression were similar at the different stages of growth, suggesting that additional factors may be involved in the clinical progression of skeletal deformities observed during growth spurts."

"Age group (days old) Body weight (g) Growth-plate thickness (μm) Static load (N) Plug area (mm2) Stress (kPa)

21 50.2 ± 5.0   741.9 ± 69.0 0.27 ± 0.03 22.4 ± 3.2 12.2 ± 1.3

35 143.3 ± 8.1 634.4 ± 76.6 0.77 ± 0.04 39.9 ± 3.9 19.5 ± 2.0

56 203.6 ± 9.4 470.9 ± 89.6 1.10 ± 0.05 44.1 ± 5.4 25.2 ± 3.4

80 251.8 ± 20.0 207.5 ± 20.5 1.36 ± 0.11 46.0 ± 5.2 29.8 ± 4.1"

Here's an image of the growth plates with and without load:

"load did not appreciably perturb the basic metabolism of chondrocytes and cell viability was preserved"

A study in vivo of the effects of a static compressive load on the proximal tibial physis in rabbits.

"Static compressive loads (10 N or 30 N) were applied for two or six weeks across one hind limb proximal tibial physis of thirteen-week-old female New Zealand White rabbits (n = 18). The contralateral hind limb in all rabbits underwent sham surgery with no load to serve as an internal control. Harvested physes were divided into portions.

Compared with unloaded shams, physes loaded at 10 N or 30 N for two weeks and at 10 N for six weeks showed histological changes in cells and matrices. Physes loaded at 30 N for six weeks were decreased in thickness and had structurally disorganized chondrocyte columns, a decreased extracellular matrix, and less intense type-II and X collagen immunohistochemical staining. Quantitative reverse transcription-polymerase chain reaction analysis of loaded samples compared with unloaded shams yielded a significantly decreased gene expression of aggrecan and type-II and X collagen and no significant changes in the matrix metalloprotease-13 gene expression with increasing load.

Compressed rabbit physes generate biochemical changes in collagens, proteoglycan, and cellular and tissue matrix architecture. Changes potentially weaken overall physeal strength, consistent with the Hueter-Volkmann principle, and lend understanding of the causes of pathological conditions of the physis."

"Compared with unloaded shams, six-week, 30-N-loaded physes were decreased in thickness and stained less intensely for collagen. The 30-N specimens also had structurally altered cells, lacunae, and chondrocyte columns (chondrons) as well as decreased extracellular matrix areas between cells and chondrons in proliferating and hypertrophic tissue zones. On enlargement compared with unloaded shams, structural changes in 30-N physes were marked by apparently fewer chondrocytes in all physeal regions, enlarged cell lacunae, irregularly aligned chondrons shortened in height, and disorganized cell arrangements within chondrons. The latter included individual chondrocytes in 30-N samples that varied in size and shape and failed to stack in an orderly, linear manner, one on the other, as did chondrons of unloaded shams"

This provides evidence that the growth plates physically push the bone apart as they are affected by load on it.  Although epiphyseal distraction helps the bones push apart and does not result in adult height gain.  Although epiphyseal distraction cannot help the growth plates with the ossification stage.

Chronic in vivo load alteration induces degenerative changes in the rat tibiofemoral joint.

"A varus loading device was attached to the left hind limb of mature rats to apply increased compression to the medial compartment and decreased compression to the lateral compartment of the tibiofemoral joint of either 0% or 100% body weight for 0, 6 or 20 weeks.

Increased compression in the medial compartment produced significant degenerative changes consistent with the development of osteoarthritis (OA) including a progressive decrease in cartilage aggregate modulus (43% and 77% at 6 and 20 weeks), diminished cellularity (38% and 51% at 6 and 20 weeks), and increased histological degeneration. At 20 weeks, medial compartment articular cartilage thickness decreased 30% while subchondral bone thickness increased 32%{consistent with endochondral ossification} and subchondral bone modulus increased 99%. Decreased compression in the lateral compartment increased calcified cartilage thickness, diminished region-specific subchondral bone thickness and revealed trends for reduced cellularity and decreased articular cartilage thickness at 20 weeks."

"Chronic increased load of 44% BW applied to the rabbit knee for 12 h/day over 12 weeks resulted in increased articular cartilage thickness and permeability with minimal fibrillation of the articular surfaces"

"9-month-old, male, Sprague–Dawley rats (weight: 666 ± 32 g)"


Strain-dependent recovery behavior of single chondrocytes.

"In this study, the compressibility and recovery behaviors of single chondrocytes were determined as a function of compressive strains from 6 to 63%. Bovine articular chondrocytes from the middle and deep zones were subjected to this range of strains, and digital videocapture was used to track changes in cell dimensions during and after compression. The normalized volume change, apparent Poisson's ratio, residual strain after recovery, cell volume fraction after recovery, and characteristic recovery time constant were analyzed with respect to axial strain. Normalized volume change varied as a function of strain, demonstrating that chondrocytes exhibited compressibility. The mean Poisson's ratio[the negative ratio of transerse to axial strain] of chondrocytes was found to be 0.29 +/- 0.14, and did not vary with axial strain{a value below 0.5 means the cell is compressible}. In contrast, residual strain, recovered volume fraction, and recovery time constant all depended on axial strain. The dependence of residual strain and recovered volume fraction on axial strain showed a change in behavior around 25-30% strain, opening up the possibility that this range of strains represents a critical value for chondrocytes."

"The average height and diameter of chondrocytes were found to be 8.4 ± 1.8μm and 11.1 ± 0.8μm, respectively. Chondrocytes were either elliptical (n = 16) or hemi-elliptical (n = 18) in appearance. All parameters determined in this study were insensitive to cell shape. The average cell volume was calculated to be 430 ± 251μm^3."

"the recovery behavior of single chondrocytes, but not their compressibility, depended on axial strain."

"At low strain levels (< 20%), cells recovered relatively quickly, and recovery was generally over 90% of the cell’s original height. At intermediate strains (20–30%), some cells exhibited similar recovery behavior, while others did not recover as quickly and showed significant residual strain"

"Above [25-30%] strain level, chondrocytes showed an impaired ability to recover, as evidenced by increased residual strain and decreased volume fraction after recovery, which differed significantly from chondrocytes compressed less than 25%. Below 25% axial strain, residual strain and recovered volume fraction showed weaker or non-existent dependence on axial strain. This suggests that 25–30% compressive strain may represent a threshold akin to the yield strain, above which some plastic deformation may occur."<-Maybe above 30% tensile strain produces similar changes.  Are these plastic changes good or bad for growth.

"Chondrocytes decreased in volume with increasing axial strain"<-could they increase in volume with increasing tensile strain?

"Though the individual components that comprise the cell, e.g., cytoskeletal filaments, cytoplasm, and plasma membrane, may be intrinsically incompressible, fluid exudation through the plasma membrane would result in a net volume loss."

It is important to note that these chondrocytes were not in the ECM.

The effects of mechanical loading on the mRNA expression of growth-plate cells.

"This in vitro study investigated the effects of mechanical loading on the mRNA expression pattern of key molecular components of the growth-plate. Short-term static loading was applied to rat proximal tibial growth-plate explants. Various age groups at specific developmental stages were investigated. In situ hybridization was used to assess the mRNA expression of the cells in different zones of the growth-plate. Four key components were investigated: 18s (basic cell metabolism), type II collagen (major extracellular matrix component), type X collagen (matrix component in hypertrophic zone) and PTH-PTHrP receptors (pre-hypertrophic chondrocytes). The spatial variation in the mRNA expression between loaded explants and their contralateral controls was compared to establish: -the sensitivity of the different growth-plate zones to mechanical loading; -the sensitivity of the different developmental stages to loading. Preliminary results indicated that static loading on the growth plate of 80 d.o. rats affects type II and X collagen gene expressions while PTH-PTHrP remains insensitive to static loading."

Couldn't get full study.

Rbp4

Retinol-binding protein 4 is expressed in chondrocytes of developing mouse long bones: implications for a local role in formation of the secondary ossification center.

"Retinol-binding protein 4 (Rbp4) is the major carrier of retinol in the bloodstream, a retinoid whose metabolites influence osteogenesis, chondrogenesis and adipogenesis. Rbp4 is mainly produced in the liver where it mobilizes hepatic retinol stores to supply other tissues. Rbp4 was present in a variety of locations in developing embryonic and postnatal mouse hindlimbs. Rbp4 was present in a restricted population of epiphyseal chondrocytes and perichondral cells correlating to the future region of secondary ossification. With the onset of secondary ossification, Rbp4 was detected in chondrocytes of the resting zone and in chondrocytes that bordered invading cartilage canals and the expanding front of ossification. Rbp4 was less abundant in proliferating chondrocytes involved in primary ossification."

"Serum retinol, complexed to Rbp4, is taken up into cells via the Rbp4 receptor, Stra6. In the cell, retinol metabolites (collectively known as retinoids) can interact with retinoic acid/rexinoid receptor (RAR/RXR) heterodimers to activate gene transcription. Abnormal retinoid levels impact bone growth"

"Rbp4 was detected in chondrocytes toward the middle of the epiphysis, in the region where the secondary ossification center will form"

"ucleated cells with small amounts of cytoplasm did not contain Rbp4 whereas Rbp4 was abundant in the cytoplasm where the nucleus had begun to deteriorate and was still present in cells without nuclei. With complete breakdown of the cell, Rbp4 was no longer detected in empty lacunae"

"Rbp4 is localized to chondrocytic cells in three regions of embryonic tibia; two relating to secondary ossification and the third relating to primary ossification. Primary ossification contributes to elongation of the limbs, whereas secondary ossification mediates expansion and mineralization of the epiphyses. Formation of the secondary ossification center is preceded by invagination of the perichondrium and invasion of the epiphyseal cartilage by vascularised canals"

Link N

The effect of Link N on differentiation of human bone marrow-derived mesenchymal stem cells.

"MSCs isolated from the bone marrow of three osteoarthritic patients{the gene expression of OA MSCs is different but it usually favors osteogenic rather than chondrogenic differentiation} were cultured in chondrogenic or osteogenic differentiation medium without or with Link N for 21 days.

Link N alone did not promote MSC chondrogenesis. However, after MSCs were supplemented with Link N in chondrogenic differentiation medium the quantity of GAG secreted into the culture medium, as well as aggrecan, COL2A1 and SOX9 gene expression, increased significantly. The gene expression of COL10A1 and osteocalcin (OC) were down-regulated significantly. When MSCs were cultured in osteogenic differentiation medium, Link N supplementation led to a significant decrease in mineral deposition, and alkaline phosphatase (ALP), OC and RUNX2 gene expression.

Link N can enhance chondrogenic differentiation and down-regulate hypertrophic and osteogenic differentiation of human MSCs."

"Link N (DHLSDNYTLDHDRAIH) is the N-terminal peptide of link protein, a glycoprotein that stabilizes the non-covalent interaction between an aggrecan G1 domain and hyaluronate"

Link N stimulates Col2a1 and decreases Col10a1 in normal human MSCs.

"MSCs were obtained from aspirates of the intramedullary canal of three osteoarthritic patients (40-60 years of age) undergoing total hip replacement"

"Since the DNA content of the cultures did not change significantly between days 3 and 21 of culture, the enhanced GAG synthesis is likely the result of increased production by each cell rather than a consequence of more cells due to cell proliferation"

"Link N alone does not directly stimulate chondrogenesis in a manner analogous to TGFβ, but rather enhances ongoing chondrogenesis"

"The ability of Link N to decrease the expression of ALP and OC may be through down-regulating the transcription regulator RUNX2"


The Effect of Link N on the Differentiation of Human Mesenchymal Stem Cells

"MSCs were isolated from the bone marrow of osteoarthritis (OA) patients. The cells were cultured in 24-well plates (3000 cells/well) in chondrogenesis differentiation medium (Invitrogen, Canada) according to the manufacturer's instructions. Link N was dissolved in the media with a final concentration of 0.1 µg/mL and 1 µg/mL, respectively. Medium without Link N was applied as a control. The media were changed every 3 days, and the used media were collected and stored at −20°C for GAG analysis. For gene expression analysis, the cells were cultured for 7, 14, and 21 days.

With the concentrations of 0.1 and 1.0 µg/mL, no toxic effect of Link N on MSCs was observed. After MSCs were cultured for 7 days and 14 days, the expression of aggrecan (ACAN), collagen II (COL2A1) and the transcription regulator SOX9 increased significantly in the media with 0.1 µg/mL or 1.0 µg/mL Link N. No significant differences were observed at day 21. When cells were cultured with 0.1 or 1.0 µg/mL Link N, the quantity of GAG in the media increased significantly at day 9, 12, and 15, but no major difference was observed at day 3 and 6 and at day 18 and 21. When MSCs were cultured for 7, 14, and 21 days, no significant effect of Link N on the expression of alkaline phosphatase (ALP) was observed. Link N significantly inhibited osteocalcin (OC) gene expression after culturing MSCs for 21 days."

We need to find some studies on Link N on normal human bone marrow.

Grow Taller by Delaying Puberty?

The effects of delayed puberty on the growth plate.

"Many athletes are beginning intense training before puberty, a time of increased bone accrual when up to 25% of total bone mineral accrual occurs. Female athletes experiencing late or delayed pubertal onset may have open epiphyseal plates that are vulnerable to injury. This investigation's purpose was to determine whether a delay in puberty (primary amenorrhea) affects the growth plate immediately postpuberty and at maturity.
Forty-eight female Sprague-Dawley rats (23 d old) were randomly assigned to 4 groups (n=12); short-term control (C-ST), long-term control (C-LT), short-term GnRH antagonist (G-ST), and long-term GnRH antagonist (G-LT). At 25 days of age, daily gonadotropin-releasing hormone antagonist (GnRH-a) injections were administered delaying pubertal onset. Left tibias were analyzed. Stained frontal slices of proximal tibia (5 µm thick) were analyzed in hypertrophic, proliferative, and reserve zones for total height, zone height, and cell/column counts.
Growth plate height was 19.7% wider in delayed puberty (G-ST) group and at maturity was 27.9% greater in G-LT group compared with control (C-LT). No significant differences were found in short-term or long-term growth plate zone heights or cell/column counts between groups. Growth plate zone height normalized to total height resulted in 28.7% larger reserve zone in the short-term GnRH-a group but the proliferative zone was 8.5% larger in the long-term group compared with the control group. Normalized to growth plate height a significant decrease was found in column counts in proliferative zones of the short-term and long-term GnRH-a groups.
Current data illustrate that delayed puberty using GnRH-a injections results in significant growth plate height and decreases proliferative column counts and zone height, thus potentially contributing to decreases in bone mass at maturity.
Growth plate height increases indicate increased potential for growth and bone accrual. However, previous models report decreased bone volume following delayed puberty via GnRH-a injections that may have detrimental effects in the long term."

"Estrogen in low doses stimulates growth hormone and results in the chondroblast progenitor cells in the reserve zone beginning clonal expansion. Suppressed estrogen levels during delayed puberty and secondary amenorrhea may have a direct negative effect on the growth plate and thus bone development"

"The GnRH-a model significantly delays the onset of puberty resulting in suppressed estradiol levels during growth"

"proximal growth plates of the tibia of IGF-1-deficient and acid-labile subunit knockout mice (LID+ALSKO) were smaller in total height and in the height of the proliferative and hypertrophic zones of chondrocytes compared with controls"

"delayed pubertal onset in the GnRH-a groups resulted in increased height[growth plate height not body height] but bone bridging would indicate that even though statistically growth plates are at different heights senescence was near with growth plate fusion at a time point similar to control animals. Delayed growth plate senescence following the dexamethasone injections was indicated by the lack of growth plate fusion in the experimental animals. After the 16-week period, the control group had 74% more fused growth plates in comparison with the dexamethasone group."

According to the author, there was no statistically significant differences in bone length between either the long term or short term groups.

"Means were very similar between groups at both time points."

Benefit of postponing normal puberty for improving final height

"In children with central precocious puberty, a GnRH analog (GnRHa) alone is efficacious in increasing final height, but in other conditions a combination of growth hormone (GH) and GnRHa is needed. In GH-deficient children with early onset of puberty and poor height prediction, the combination of GH and GnRHa increases final height by 1.0-1.3 s.d. In children with idiopathic short stature and persistent short stature after intrauterine growth retardation, the combination also appears to be beneficial. Potential side effects include weight gain, a negative effect on bone mineralization, and psychosocial consequences. More data on long-term safety have to be collected before the combination of GH and GnRHa in children with idiopathic short stature should be considered for clinical use outside clinical trials. "

"within the normal range, late developers have a greater leg length:sitting height ratio than early developers"

Syndecan 4

Syndecan 4 is upregulated by LSJL.


Bone fracture repair, but not fetal skeletal development is supported by syndecan-4.

"We used Sdc4-/- mice to analyze the functional role of Scd4 in endochondral ossification of mouse embryos and in adult fracture repair. In Sdc4-/-/LacZ knock-in animals, Sdc4 promoter activity was detectable in all stages of chondrocyte differentiation, and Sdc4 deficiency inhibited chondrocyte proliferation. Aggrecan turnover in the uncalcified cartilage of the epiphysis was decreased transiently in vivo, but this did not lead to a growth phenotype at birth. By contrast, fracture healing in adult mice was markedly delayed in Sdc4-/- animals and accompanied by increased callus formation. Blocking inflammation during fracture healing with a TNFα inhibitor reduced these changes in Sdc4-/- animals to WT levels. Analysing the discrepancy between the mild embryonic and the severe adult phenotype, we found a compensatory up-regulation of Sdc2{down} in the developing cartilage of Sdc4-/- mice that was absent in adult tissue. Stimulation of chondrocytes with Wnt3a in vitro, led to an increased expression of Sdc2, while stimulation with TNFα resulted in an up-regulation of Sdc4 but a decreased expression of Sdc2. TNFα stimulation decreased Sdc2- and increased Sdc-4 expression even in presence of Wnt3a, suggesting a strong effect of inflammation on the regulation of Sdc expression."

The upregulation of Sdc4 and downregulation of Sdc2 is consistent with the inflammatory role in fracture repair thus providing evidence that inflammation is involved in the LSJL response.

"Expression of Sdc1, -2 and -4 has been detected in chondrocytes and progenitor cells during development of rat mandibular condyles. Sdc3 has been implicated in regulating the size of skeletogenic condensations and growth factor-mediated proliferation of chondrocytes during limb development and growth. In addition, Sdc2 and -4 are involved in osteoblast cell adhesion and survival"

"At the cellular level, Sdc4 promoter activity was seen throughout the cartilage of the epiphysis and in resting, proliferating, prehypertrophic and hypertrophic chondrocytes within the epiphyses"

"a substantially reduced staining for ADAMTS-4{up in LSJL} protein was detected throughout the cartilage in Sdc4-/- E16.5 tibia in comparison with wild type cartilage"

"Sdc2 expression during development is most likely induced by Wnt3a, a growth factor that is involved in the transition of mesenchymal cells into chondroprogenitor cells"

"stimulation of WT chondrocytes with 100 ng/ml Wnt3a in vitro led to increased Sdc2 mRNA levels (3 fold up-regulation). In contrast TNFα stimulation decreased Sdc2 mRNA levels in a dose dependent manner by up to 50 %"

Sdc4 and Sdc2 regulation are likely highly involved in the LSJL response.

Sdc2 mRNA levels in a dose dependent manner by up to 50 %"

reseveratrol

Biological effects of the plant-derived polyphenol resveratrol in human articular cartilage and chondrosarcoma cells.

"RSV is chondroprotective for articular cartilage in rabbit models for arthritis{Can this translate to height benefits?}. Effects of RSV on human articular cartilage homeostasis were studied by assessing production of matrix-degrading enzymes (MMP-13, ADAMTS4, and ADAMTS5), as well as proteoglycan production and synthesis. The counteractions of RSV against catabolic factors (e.g., FGF-2 or IL-1β) were examined by in vitro and ex vivo using monolayer, three-dimensional alginate beads and cartilage explants cultures, respectively. RSV improves cell viability of articular chondrocytes and effectively antagonizes cartilage-degrading protease production that was initiated by catabolic and/or anti-anabolic cytokines in human articular chondrocytes. RSV significantly enhances BMP7-promoted proteoglycan synthesis. RSV inhibits the activation of transcription factors involved in inflammation and cartilage catabolic signaling pathways, including direct downstream regulators of MAPK (e.g., AP-1, PEA3) and NFκB. RSV selectively compromises survival of human chondrosarcoma cells, but not primary articular chondrocytes, revealing cell-specific activity of RSV on non-tumorigenic versus tumor-derived cells. RSV exerts its chondroprotective functions, in part, by deactivating p53-induced apoptosis in human primary chondrocytes, but not human chondrosarcoma."

"signaling cascades generated by inflammatory cytokines (e.g., IL-1) or fibroblast growth factor-2 (FGF-2 or basic FGF) favor catabolism by stimulating protease production and inhibiting proteoglycan deposition in human adult articular cartilage or intervertebral disc tissue via ERK/MAPK activation"

"FGF-2 mediates striking antagonistic effects on cartilage anabolic activity in conjunction with IGF-1 and BMP7, and both FGF-2 and IL-1 modify chondrocyte gene expression when stimulated by mechanical injury "

"p53 DNA binding activity is significantly stimulated by IL-1β (10 ng/ml) along with AP-1, AP-2, Ets1/PEA3, NFκB, p53, Sp1, and multiple STATs that are critical for cytokine signals"

GPR177

Gpr177, a novel locus for bone-mineral-density and osteoporosis, regulates osteogenesis and chondrogenesis in skeletal development.

"Gpr177 exhibits an ability to modulate the trafficking of Wnt similar to the Drosophila Wls/Evi/Srt.  To overcome the early lethality associated with the inactivation of Gpr177 in mice, conditional gene deletion is utilized to assess its functionality. Here we report the generation of four different mouse models with Gpr177 deficiency in various skeletogenic cell types. The loss of Gpr177 severely impairs development of the craniofacial and body skeletons, demonstrating its requirement for intramembranous and endochondral ossifications, respectively. Defects in the expansion of skeletal precursors and their differentiation into osteoblasts and chondrocytes suggest that Wnt production and signaling mediated by Gpr177 cannot be substituted. Because the Gpr177 ablation impairs the secretion of Wnt proteins, we therefore identify their sources essential for osteogenesis and chondrogenesis. The intercross of Wnt signaling between distinct cell types is carefully orchestrated and necessary for skeletogenesis.  Gpr177 controls skeletal development through modulation of autocrine and paracrine Wnt signals in a lineage-specific fashion"

"formation of the calvarial, maxillary and mandibular bones mediated by intramembranous ossification is defective or completely missing in the Gpr177Dermo1 embryos"

Loss of GPR177 delays chondrocyte hypertrophy.

"The chondrogenic deletion of Gpr177 significantly reduced bone mineralization, and interfered with chondrocyte maturation"

"The number of cells undergoing mitotic division is significantly reduced in the columnar zone, but not epiphyses of the Gpr177Dermo1 and Gpr177Col2 humeruses, suggesting that expansion of the proliferating and prehypertrophic but not the resting chondrocytes was affected by the loss of Gpr177"

"disruption of endochondral ossification starts at chondrocyte maturation, and the subsequent events, including ECM remodeling, vascular invasion and osteoblastogenesis, are impaired in the Gpr177Dermo1 mutants."  Chondrocyte maturation is where Beta Catenin begins to take over for Sox9.

"Gpr177 is dispensable in the osteoblasts during intramembranous and endochondral ossifications. Our findings suggest that the impairment of osteoblast differentiation in the Gpr177Dermo1 and Gpr177Col2 limbs is attributed to delay in chondrocyte maturation but not intrinsic defects of the osteoblasts."

Adipose Tissue Stem Cells to Chondrocytes

High plasticity of pediatric adipose tissue-derived stem cells: too much for selective skeletogenic differentiation?

"We have used different methods to establish stem cells from adipose tissue (adipose-derived stem cells [ADSCs], adipose explant dedifferentiated stem cells [AEDSCs]) from several pediatric patients and investigated their phenotype and differentiation potential using monolayer and micromass cultures. We have also addressed the overlooked issue of selective induction of cartilage differentiation. ADSCs/AEDSCs from different patients showed a remarkably similar behavior. Pluripotency markers were detected in these cells, consistent with ease of reprogramming to induced pluripotent stem cells. Significantly, most ADSCs expressed markers of tissue-specific commitment/differentiation, including skeletogenic and neural markers, while maintaining a proliferative, undifferentiated morphology. Exposure to chondrogenic, osteogenic, adipogenic, or neurogenic conditions resulted in morphological differentiation and tissue-specific marker upregulation. These findings suggest that the ADSC "lineage-mixed" phenotype underlies their significant plasticity, which is much higher than that of chondroblasts we studied in parallel. Finally, whereas selective ADSC osteogenic differentiation was observed, chondrogenic induction always resulted in both cartilage and bone formation when a commercial chondrogenic medium was used; however, chondrogenic induction with a transforming growth factor β1-containing medium selectively resulted in cartilage formation."

"ADSCs expressed the mesenchymal markers CD44 (hyaluronic acid receptor), CD90 (Thy-1), and CD105 (endoglin)"

" with the exception of SOX2, transcripts for the pluripotency markers c-MYC, OCT4, NANOG, KLF4, and DNMT3B were detected in ADSCs from all patients tested"  Surprisingly, ADSC's do not express type II collagen by default.  They do express Aggrecan.  Type II collagen was induced by TGF-Beta1.

Chondroblasts in contrast to ADSC's express ColII and ALP by default.