Thyroid-Stimulating Hormone Favors Runx2-Mediated Matrix Mineralization in HOS and SaOS2 Cells: An In Vitro and In Silico Approach

[helvella]

A recent paper which discusses the very detailed effects of TSH. Like very other paper, just a step on the way - but interesting.

This sentence from the Discussion was interesting:

These findings clearly indicate that TSH acts in a similar fashion in thyrocytes as well as in osteoblasts. If so, it is essential to know the mechanism of TSH action in this regard.

Molecules.2022 Feb; 27(3): 613.

Published online 2022 Jan 18. doi: 10.3390/molecules27030613

PMCID: PMC8838199

PMID: 35163879

Thyroid-Stimulating Hormone Favors Runx2-Mediated Matrix Mineralization in HOS and SaOS2 Cells: An In Vitro and In Silico Approach

Ramajayam Govindan,1,2,*† Mohamed El-Sherbiny,3 Khalid Mohamed Morsy Ibraheem,4 Srinivasan Narasimhan,2 Mohamed EL-Dosoky Mohamed Salama,5 Fazil Ahmad,4 Selvaraj Jayaraman,6,*† Vishnu Priya Veeraraghavan,6 Srinivasan Vengadassalapathy,7 Surapaneni Krishna Mohan,8 Vidhya Rekha Umapathy,9 Gayathri Rengasamy,6 Shazia Fathima Jaffer Hussain,10 Maheshkumar Poomarimuthu,1 and Senthilkumar Kalimuthu11

Muralikannan Maruthamuthu, Academic Editor, Nagendran Tharmalingam, Academic Editor, and Aruliah Rajasekar, Academic Editor

Abstract

Osteoporosis is a skeletal disease that is both systemic and silent characterized by an unbalanced activity of bone remodeling leading to bone loss. Rising evidences demonstrate that thyroid stimulating hormone (TSH) has an important role in the regulation on the metabolism of bone. However, TSH regulation on human osteoblast essential transcriptional factors has not been identified. Current study examined the role of TSH on human osteoblastic Runx2 expression and their functional genes by in vitro and in slico analysis. Human osteoblast like (HOS and SaoS-2) cells were cultured with DMEM and treated with hTSH at the concentration of 0.01 ng/mL and 10 ng/mL. After treatment, osteoblastic Runx2 and IGF-1R beta expression were studied using RT-PCR and western blot analysis. TSH treatment induced osteoblastic essential transcriptional factor, Runx2 in HOS and SaOS2 cells on 48 h duration and elevated the expression of IGF-IR β gene and Protein in SaoS-2 cells. TSH also promotes Runx2 responsive genes such as ALP, Collagen and osteocalcin in SaOS2 cells on day 2 to day 14 of 10 ng/mL of treatment and favors’ matrix mineralization matrix in these cells. In addition, TSH facilitated human osteoblastic cells to mineralize their matrix confirmed by day 21 of alizarin red calcium staining. In silico study was performed to check CREB and ELK1 interaction with Runx2. Results of in silico analysis showed that TSH mediated signalling molecules such as CREB and ELK1 showed interaction with Runx2 which involve in osteobalstic gene expression and differentiation. Present findings confirm that TSH promotes Runx2 expression, osteoblastic responsive genes and bone matrix formation.

Keywords: TSH, Runx2, ALP, SaOS2, CREB, ELK1

Full paper freely accessible here:

ncbi.nlm.nih.gov/pmc/articl...

jimh111

[jimh111]

TSH seems to have multiple functions. I don't understand all this stuff but it seems obvious that ideally we all need TSH. This presents problems for those of us with subnormal TSH secretion. Not only do we lack essential TSH with loss of effects on deiodinase but if there are bone problems the patient and thyroid therapy is blamed.

By the way I think hTSH refers to human recombinant TSH which is a manufactured form of TSH. Since TSH has many isoforms I'm not sure we can always rely on experiments with hTSH. I struggle with this stuff so don't take this as Gospel.

[TSH110 in reply to jimh111]

Would it be causing my myriad of bone problems and tendonopathies? I have had some decades before my atropic autoimmune thyroiditis was diagnosed. Things got worse on Levothyroxine and have not be fully reversed by NDT. I feel like pants if my TSH is not very low/suppressed

[jimh111 in reply to TSH110]

To be honest I don't know.

I have had a TSH near zero for over two decades with no bone problems. I have also heard of many other similar cases so I just assume the role of TSH in bone formation is minor. It could be more or less important according to our individual genetics or perhaps it has its main role during early development.

To sum up the experiments find a TSH effect on bone but I've never seen any study in people or animals.

[TSH110 in reply to jimh111]

I just thought it was interesting and wondered, as no one really seems to know quite why musculo-skeletal problems are more prevalent in those with thyroid disorder (even when treated). I’m sure genetics must play an important role.

[jimh111 in reply to TSH110]

I think a lot of the muscular problems are due to inadequate treatment of hypothyroidism causing muscle problems and enhanced sensation of pain. Some of the skeletal problems could be similar, minor skeletal malformations that would not cause problems in an euthyroid person.

[TSH110 in reply to jimh111]

I get the feeling with mine that the healing process of minor trauma is not functioning as efficiently as it ought to. Even in my early 30’s all my upper vertebrae had extra bone and spurs hence the Ankylosing spondilitis diagnosis I went because I had been plagued with trapped nerves affecting either legs or arms, one at a time, since my early 20’s. I went to an osteopath - doc said it was a good idea - who worked wonders on it. Now I wake with horrible back ache and have trouble getting going but it eases after an hour or so. I get bouts of it then nothing for ages. Trapped nerves have stopped (touch wood). Doc said the extra bone was causing the nerves to get caught. I suppose the bone modelling mechanism has gone wrong with too much making and not enough modelling going on. The Achilles tendinitis involved lumps forming in the tendon on the first one and I was told it might be calcificationof the tendon which is something boney appearing where it shouldn’t. I ended up needing a decompression. 30 years later the other one is doing the same ☹️ I don’t fancy the op it wasn’t very nice but just getting on an interminable list for one, these days, sounds highly unlikely especially once you’re over the 60 hill you have to carry on working and paying NI but you aren’t worth treating if anything goes wrong…. Then there’s the bakers cysts……they don’t bother me now they are quite big 🤣🤣🤣

[BB001 in reply to TSH110]

I have had suppressed TSH for 30 years and no bone problems.I have atrophic thyroiditis too, so like you have lost my thyroid's ability to both produce T3 and to convert T4 to T3

Thus I think it's more likely that low thyroid hormones T4 and T3 are responsible for your bone problems, and TSH is just a bystander.

[TSH110 in reply to BB001]

But I feel very well bar these problems and my last tests showed optimal levels of free t3 (I take NDT) it’s as if something is putting those particular things out of whack. I know the problems are more common in those with thyroid disorder.

[greygoose in reply to TSH110]

Tests can't show optimal T3 because optimal is not a number, it's the way you feel. If you don't feel optimal, then you aren't.

[TSH110 in reply to greygoose]

Well in the top 1/3 but the blood test was a while back now. I do just go on how I feel - mentally very good indeed, especially compared to the chronic and debilitating depression I suffered from before taking NDT, that alone is worth it’s weight in gold. I feel fine, bar the arthritis/ankylosing spondilitis/Achilles tendinitis - must be my heel. Also it could be decades of manual work taking its toll on me.

[greygoose in reply to TSH110]

Perhaps it needs to be higher than the top third. Or perhaps the level has dropped since. But good that the depression has gone.

[TSH110 in reply to greygoose]

I wonder if it’s a vitamin/mineral imbalance. My iron was not as good as it could be on that last test but the rest seemed decent. I take betteryou iron and vitd with k2 a few times a week but not every day and b complex once a week. I get a lot of cramp in my feet and calves and now my hands but it’s intermittent and I see no pattern to it at all, except stirring things in pots on the cooker seems to set the hand cramp off. I suspect I have dupchrens in one hand but the cramp is always in both at once, they look really bizarre too fingers all over the place. They are all irritating but not very debilitating. Would be interesting to get to the bottom of them tho.

Yes wonderful to have got rid of that awful depression, I don’t know how I kept going being as miserable as that for decades!

I could try upping the NDT but I just get a feeling it’s right at the moment.

[greygoose in reply to TSH110]

Do you also take magnesium? Low magnesium causes cramps.

[TSH110 in reply to greygoose]

I eat a lot almonds, some every day, and pop the occasional magnesium malate tablet. I don’t think it’s magnesium per say but perhaps it’s due to an electrolyte imbalance with that and calcium and maybe potassium is one of them? (I eat a daily banana and sweet potatoes about once a week which are supposed to be good in that) I don’t take any calcium supplements. If it gets really bad I resort to quinine, which usually stops it. It’s a mystery to me what’s causing it. I used to take selenium but was a bit trepidaceous about it and stopped it after a month ditto with zinc.