Simulated Microgravity Reduces Growth but Preserves Bone-Relevant Bioactivity of Medicago truncatula

Gary W. Stutte; Adam Shinners

doi:10.2478/gsr-2026-0007

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Simulated Microgravity Reduces Growth but Preserves Bone-Relevant Bioactivity of Medicago truncatula

Gravitational and Space Research

Volume 14 (2026): Issue 1 (January 2026)

By: Gary W. Stutte and Adam Shinners

Open Access

|Jun 2026

Figures & Tables

Two-dimensional clinostat system positioned with a controlled environment chamber (EGC M-48, Environmental Growth Chamber, Chagrin Falls, OH, USA). M. truncatula seeds were placed at the center of each petri dish containing Murashge and Skoog (MS) medium and mounted on the rotating clinostat plate to simulate microgravity conditions, while identical controls were maintained within the same chamber (foreground) under identical environmental conditions.

Effects of simulated microgravity on growth of M. truncatula growth parameters after 14 days. The data represent (A) shoot fresh mass, (B) shoot length, (C) primary root length, and (D) number of lateral roots under stationary 1g or clinorotation (simulated μg) conditions. Data represent the mean of three independent experiments (10 seedlings per experiment). Error bars indicate standard deviation. Asterisks denote statistically significant differences (p ≤ 0.001) between 1g and simulated μg treatments.

Effects of simulated microgravity on M. truncatula secondary metabolite concentrations after 14 days. The data represents (A) total flavonoid concentration (mg g−1 fresh mass) as quercetin equivalents and (B) total isoflavonoid concentration (mg g−1 fresh mass) as genistein equivalents. Data represent the mean of three independent experiments (10 seedlings per experiment). Error bars indicate standard deviation. Asterisks denote statistically significant differences (p ≤ 0.05) between 1g and simulated μg treatments.

RAW264.7 activity in response to the 0.1% ethanol extracts from 14-day-old M. truncatula seedlings that have been grown under static or 2-D clinostat conditions.

Representative morphology of M. truncatula seedlings grown for 14 days under (A) stationary 1g conditions and (B) 2-D clinostat-modeled simulated microgravity. Seedlings grown under microgravity exhibited reduced shoot length, primary root length, lateral root development and overall biomass reduction relative to 1g controls.

RAW264_7 metabolic activity expressed as percentage of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

Treatment
	Static			Clinostat
Ethanol controls
Dose (%)	% Control	SD	Dose (%)	% Control	SD
0.1	100	3.7	0.1	100	3.8
M. truncatula extracts
	Static			Clinostat
Dose (ng)^*	% Control	SD	Dose (ng)	% Control	SD
4.9	58	1.4	5.4	56.6	1.2
9.8	53.2	3.5	10.7	54.3	2.6
14.7	52.3	2.6	16.5	51.8	7.2
19	51.8	4.4	21.4	40.2	1.6

MC3T3-E1 metabolic activity expressed as percent of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

Treatment

	Static			Clinostat
Ethanol controls
Dose (%)	% Control	SD	Dose (%)	% Control	SD
0.1	100	7.8	0.1	100	8.3
M. truncatula extracts
	Static			Clinostat
Dose (ng)^*	% Control	SD	Dose (ng)	% Control	SD
4.9	86.1	4.2	5.4	97.8	2.5
9.8	94.4	8.3	10.7	97.2	2.8
14.7	97.2	6.9	16.5	96.7	8.9
19	95.8	8.3	21.4	95.8	8.3

Alkaline phosphatase (ALP) metabolic activity expressed as percent of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

Treatment

	Static			Clinostat
Ethanol controls
Dose (%)	% Control	SD	Dose (%)	% Control	SD
0.1	100	7.2	0.1	100	7.3
M. truncatula extracts
	Static			Clinostat
Dose (ng)^*	% Control	SD	Dose (ng)	% Control	SD
4.9	96.4	7.5	5.4	96.8	7.6
9.8	98.2	7.4	10.7	96.8	7.4
14.7	96.8	7.5	16.5	98.2	7.5
19	97.3	7.4	21.4	95.5	7.3

Summary of metabolic effects of Medicago truncatula ethanol extracts on bone-relevant cell models_

Cell Line	Cell type/function	End point assessed	Observed effect^x
MC3T3-E1	osteoblast-like	Cell Proliferation (cell counts)	No detectable effect
MC3T3-E1	osteoblast-like	Alkaline phosphatase activity	No detectable effect
RAW264.7	osteoclast precursor^y	Cell Proliferation (cell counts)	↓40–60 % relative to EtOH control

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DOI: https://doi.org/10.2478/gsr-2026-0007 | Journal eISSN: 2332-7774

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Language: English

Page range: 7 - 19

Published on: Jun 12, 2026

Published by: American Society for Gravitational and Space Research

In partnership with: Paradigm Publishing Services

Publication frequency: Volume open

Keywords:

simulated microgravity,

alkaline phosphatase,

plant secondary metabolites,

spaceflight biology

Related subjects:

Life sciences,

Life sciences, other,

Materials sciences,

Materials sciences, other,

Physics,

Physics, other

© 2026 Gary W. Stutte, Adam Shinners, published by American Society for Gravitational and Space Research
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 14 (2026): Issue 1 (January 2026)

Simulated Microgravity Reduces Growth but Preserves Bone-Relevant Bioactivity of Medicago truncatula

Figures & Tables

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure S1.

RAW264_7 metabolic activity expressed as percentage of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

MC3T3-E1 metabolic activity expressed as percent of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

Alkaline phosphatase (ALP) metabolic activity expressed as percent of 0_1% ethanol control following exposure to Medicago truncatula extracts under static and clinostat conditions_

Summary of metabolic effects of Medicago truncatula ethanol extracts on bone-relevant cell models_

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