Growth Hormones Exert A Direct Stimulatory Effect On Epiphyseal Cartilage And Stimulates Longitudinal Bone Growth Directly

Me: I think it is time to put the questioning of whether the hGH in the human body directly stimulated the epiphyseal plate for longitudinal growth to be answered conclusively. It does.

From article 1…

We see that chondrocytes from the tested rat’s epiphyseal plates are removed and cultured using calf serum. The thing to note is that to make the condrocyte colonies one needs the serum. There is also a strong correlation in terms that the number of colonies formed is directly proportional to the concentration of the calf serum. Overall we see that we need all three factors of

  1. Number of initial seeded chondrocytes
  2. Have enough 10-15% newborn calf serum
  3. Having around 10-40 ng/ml of hGH
They will allow for the best formation of chondrocyte colonies. The researchers conclude that “These results show that GH potentiates colony formation in chondrocytes of the epiphyseal growth plate, providing further support for the contention that GH exert a direct stimulatory effect on epiphyseal cartilage and thus stimulates longitudinal bone growth directly.” also…”The finding that GH preferentially potentiated the formation of large size colonies suggests that GH promoted the differentiation of early proliferative chondrocytes or stem cell chondrocytes with an inherent high capacity to proliferate.” This seems to suggest that the GH also has two really big functions to progenitor cells and proliferative chondrocytes, first in differentiating the progenitor cells into chondrocytes and then in helping lead to to chondrocyte proliferation.

From article 2…

We see that both hGH and IGF-1 both lead to increased cloning efficiency. What is even more interesting is that IGF-1 when used after the pretreatment of the chondrocytes by GH lead to a far greater effect of cloning efficiency. The researchers concluded that …” The results of the present study show that pretreatment of hypophysectomized rats with GH, but not with IGF-I, promotes the formation of chondrocyte colonies and make the chondrocytes susceptible to IGF-I in vitro. The results suggest that GH induces colony formation by IGF-I-independent mechanisms and that IGF-I is a second effector in GH action as previously shown

From article 3…

This study was designed to investigate whether growth hormone (GH) influences the expression of its own receptor in chondrocytes. To investigate this possibility GH-receptor mRNA was measured in cultured rat epiphyseal chondrocytes in the absence or presence of GH under various experimental conditions. The researchers concluded that “GH specifically regulates mRNA levels for its own receptor in rat epiphyseal chondrocytes by interacting with somatogenic binding sites. These findings also suggest a transcription-dependent regulatory system between the GH-receptor and the GH-receptor gene.”

Implications: It seems that GH can be used to cause cell colony formation and is the first hormone that is usually used to stimulate epiphyseal plate chondrocytes. THe IFG-1 pathway is independent of it’s pathway. It seems that the GH causes it’s own receptors to form by turning it’s own receptor genes on.

From PubMed article 1 link HERE


Endocrinology. 1986 May;118(5):1843-8.

Growth hormone potentiates colony formation of epiphyseal chondrocytes in suspension culture.

Lindahl A, Isgaard J, Nilsson A, Isaksson OG.

Abstract

The effect of human GH (hGH) on colony formation of rat epiphyseal plate chondrocytes was studied in suspension culture. Chondrocytes were isolated enzymatically from epiphyseal plates of the proximal tibia of 28-day-old normal male rats, and were cultured in a suspension stabilized with 0.5% agarose. After approximately 7 days of culture in the presence of 10% newborn calf serum (NCS), chondrocyte colonies developed consisting of varying numbers of cells in matrix. No colonies developed in the absence of NCS, and the number of formed colonies was proportional to the concentration of NCS (5-20%) in the medium. hGH potentiated the formation of large size colonies (diameter greater than 90 microns) after a culture period of 10 or 14 days. The lowest effective concentration of hGH was 10 ng/ml, while 40 ng/ml hGH gave a maximal stimulatory effect (40-50%). Higher concentrations of hGH (80 and 160 ng/ml) showed reduced potentiation of colony formation. The stimulatory effect of hGH was expressed at 10-15% of NCS at 14 days of culture. There was a linear relation between the number of seeded cells and the number of colonies formed, both in the absence and presence of hGH. These results show that GH potentiates colony formation in chondrocytes of the epiphyseal growth plate, providing further support for the contention that GH exert a direct stimulatory effect on epiphyseal cartilage and thus stimulates longitudinal bone growth directly. The finding that GH preferentially potentiated the formation of large size colonies suggests that GH promoted the differentiation of early proliferative chondrocytes or stem cell chondrocytes with an inherent high capacity to proliferate.

PMID: 3698898     [PubMed – indexed for MEDLINE]

From PubMed article 2 link HERE


Endocrinology. 1987 Sep;121(3):1070-5.

Growth hormone in vivo potentiates the stimulatory effect of insulin-like growth factor-1 in vitro on colony formation of epiphyseal chondrocytes isolated from hypophysectomized rats.

Lindahl A, Isgaard J, Isaksson OG.

Abstract

The effect of GH pretreatment in vivo on the colony formation of epiphyseal chondrocytes from hypophysectomized rats and the subsequent responsiveness to insulin-like growth factor (IGF-I) was studied in vitro. Chondrocytes from epiphyseal growth plates of the proximal tibia of 36-day-old hypophysectomized rats were enzymatically isolated and cultured in suspension, stabilized with agarose (0.5%) in Ham’s F-12 medium and serum supplement. After 14 days the cultures were terminated and screened for cloning efficiency (number of colonies with a diameter greater than 56 microns/1000 seeded cells) and for distribution of cloning efficiency as a function of colony size. Pretreatment with human GH in vivo for 24 h (10 micrograms X 3) increased the cloning efficiency during the subsequent culture period (control, 1.5 +/- 0.1; human GH, 4.4 +/- 0.3). Addition of IGF-I to the chondrocyte cultures from control rats caused a slight increase in cloning efficiency (control, 1.5 +/- 0.1; IGF-I, 2.2 +/- 0.3) but caused a marked increase in chondrocyte cultures from GH-pretreated rats (control, 4.4 +/- 0.4; IGF-I, 8.2 +/- 0.9). The cloning efficiency was increased 12 and 24 h, but not 4 h, after start of GH-treatment in vivo. The increased responsiveness to IGF-I in vivo showed a similar time course after GH pretreatment. The distribution of cloning efficiency was altered in cultures of chondrocytes isolated from the GH-pretreated rats; large colonies were overrepresented in the GH-treated group. Colonies with a diameter exceeding 180 microns were only seen in cultures of chondrocytes isolated from the GH-pretreated animals. Addition of IGF-I in vitro did not alter the distribution of cloning efficiency, but increased the mean colony size of all colonies. Pretreatment of the rats with two different doses of IGF-I in vivo for 24 h (5 micrograms X 3 or 50 micrograms X 3) had a slight stimulatory effect on subsequent colony formation, but no potentiation of IGF-I in vitro was demonstrated. The results of the present study show that pretreatment of hypophysectomized rats with GH, but not with IGF-I, promotes the formation of chondrocyte colonies and make the chondrocytes susceptible to IGF-I in vitro. The results suggest that GH induces colony formation by IGF-I-independent mechanisms and that IGF-I is a second effector in GH action as previously shown for cultured 3T3-preadipose cells.(ABSTRACT TRUNCATED AT 400 WORDS)

PMID: 3622376    [PubMed – indexed for MEDLINE]
From PubMed article 3 link HERE

Mol Cell Endocrinol. 1990 May 7;70(3):237-46.

Growth hormone regulation of the growth hormone receptor mRNA in cultured rat epiphyseal chondrocytes.

Nilsson A, Carlsson B, Mathews L, Isaksson OG.

Source

Department of Physiology, University of Göteborg, Sweden.

Abstract

This study was designed to investigate whether growth hormone (GH) influences the expression of its own receptor in chondrocytes. To investigate this possibility GH-receptor mRNA was measured in cultured rat epiphyseal chondrocytes in the absence or presence of GH under various experimental conditions. Chondrocytes were isolated enzymatically from epiphyseal growth plates of the proximal tibia of 20-day-old male rats and cultured in monolayer in Ham’s F-12 medium supplemented with 10% calf serum and 1% of a serum substitute. The cells were seeded at various densities (100,000-1,000,000 cells per flask) and cultured for 14 days. Subsequently, the calf serum-containing medium and the cells cultured for various periods of time (0-24 h) before total nucleic acid preparation. GH-receptor mRNA was measured with a solution hybridization technique using [35S]UTP-labeled RNA growth hormone receptor cloned from rat liver cDNA. Human GH (hGH; 50 ng/ml) increased GH-receptor mRNA after 3 h and maximal levels were seen 12 h after GH addition. This effect of hGH was time and dose dependent with a significant effect of hGH at a concentration of 0.5 ng/ml and a maximal effect at 50 ng/ml. The hGH-stimulated increase of GH-receptor mRNA was completely blocked by actinomycin-C1 (1.0-0.1 micrograms/ml), while cycloheximide (10 micrograms/ml) only slightly counteracted the hGH effect. Rat and human GH were equally potent, and ovine prolactin was effective at 500 ng/ml but not 5 and 50 ng/ml. A high dose of insulin-like growth factor-I (IGF-I; 1 microgram/ml) caused a small stimulatory effect and addition of 10% calf serum caused a marked increase in GH-receptor mRNA. The level of GH receptor mRNA after 14 days of culture was inversely proportional to the cell density at the start of culture. These results show that GH specifically regulates mRNA levels for its own receptor in rat epiphyseal chondrocytes by interacting with somatogenic binding sites. These findings also suggest a transcription-dependent regulatory system between the GH-receptor and the GH-receptor gene.

PMID: 1694505    [PubMed – indexed for MEDLINE]

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