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Cited by 15Bone health and osteoporosis researchBone Metabolism and DiseasesEstrogen and related hormone effects

β-Ecdysone Augments Peak Bone Mass in Mice of Both Sexes

Open access
Dai, Weiwei, Zhang, HongLiang, Zhong, Zhendong A., Jiang, Li, Chen, Haiyan, Lay, Yu-An Evan, Kot, Alexander, Ritchie, Robert O. +2 more·2015·Clinical Orthopaedics & Related Research
DOI 10.1007/s11999-015-4246-5PMC PMC4488193
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Abstract

BACKGROUND: One of the strongest predictors for osteoporosis is peak bone mass. Interventions to augment peak bone mass have yet to be developed. β-Ecdysone (βEcd), a natural steroid-like compound produced by arthropods to initiate metamorphosis, is believed to have androgenic effects and so may be used to augment bone mass. QUESTIONS/PURPOSES: The purpose of this study was to use both male and female (1) gonadal-sufficient; and (2) -insufficient mice to investigate sex differences in terms of bone development and structure after βEcd administration. METHODS: Two-month-old male and female Swiss-Webster mice were randomized to receive either vehicle or βEcd (0.5 mg/kg) for 3 weeks. In a separate experiment to evaluate the effects of βEcd on sex hormone-deficient mice, gonadectomy was performed in male (orchiectomy [ORX]) and female mice (ovariectomy [OVX]). Sham-operated and the ORX/OVX mice were then treated for 3 weeks with βEcd. Primary endpoints for the study were trabecular bone structure and bone strength. RESULTS: In male mice, the trabecular bone volume was 0.18±0.02 in the placebo-treated (PL) and 0.23±0.02 in the βEcd-treated group (p<0.05 versus PL); and 0.09±0.01 in the ORX group (p<0.05 versus PL) and 0.12±0.01 in the ORX+βEcd group. Vertebral bone strength (maximum load) was 43±2 in PL and 51±1 in the βEcd-treated group (p<0.05 versus PL); and 30±4 in the ORX group (p<0.05 versus PL) and 37±3 in the ORX+βEcd group. In female mice, trabecular bone volume was 0.23±0.02 in PL and 0.26±0.02 in the βEcd-treated group (p<0.05 versus PL); and 0.15±0.01 in the OVX group (p<0.05 versus PL) and 0.14±0.01 in the OVX+βEcd group. Maximum load of the vertebrae was 45±2 in PL and 48±4 in the βEcd-treated group; and 39±4 in the OVX group (p<0.05 versus PL) and 44±4 in the OVX+βEcd group. CONCLUSIONS: These findings suggest the potential use of βEcd in the augmentation of bone mass in growing male and female mice. It may also partially prevent the detrimental effects of gonadectomy on trabecular bone. CLINICAL RELEVANCE: Our results support the potential use of βEcd or nature products that are rich in βEcd to augment peak bone mass. βEcd may differ from the other anabolic hormone treatments that may have severe side effects such as serious cardiac complications. However, its effects on humans remain to be determined.

AI snapshot

Written from the paper’s full text.

TL;DR

In growing Swiss-Webster mice, 3-week β-ecdysone treatment increased trabecular and cortical bone formation, bone volume, and bone strength in both sexes, and partially prevented trabecular bone loss after gonadectomy. The authors conclude β-ecdysone may augment peak bone mass, but human efficacy remains untested.

Key findings

  • In intact male mice, β-ecdysone increased vertebral trabecular BV/TV from 0.18 ± 0.02 to 0.23 ± 0.02 (p < 0.05) and vertebral maximum load from 43 ± 2 to 51 ± 1 (p < 0.05); ORX reduced BV/TV to 0.09 ± 0.01 and maximum load to 30 ± 4, and ORX + β-ecdysone raised maximum load to 37 ± 3 (p < 0.05 versus ORX).
  • In intact female mice, β-ecdysone increased vertebral BV/TV from 0.23 ± 0.02 to 0.26 ± 0.02 (p < 0.05), cortical bone volume from 0.66 ± 0.1 to 0.79 ± 0.2 (p < 0.05), and femoral maximum load from 11 ± 0.5 to 12 ± 0.5 (p < 0.05); vertebral maximum load was 45 ± 2 in placebo and 48 ± 4 with β-ecdysone, without a reported p value.
  • β-ecdysone increased cortical bone formation in intact mice: periosteal BFR was 1.31 ± 0.1 to 1.80 ± 0.3 in males and 1.1 ± 0.1 to 1.7 ± 0.2 in females; endocortical BFR was 0.9 ± 0.1 to 2.6 ± 0.3 in males and 1.4 ± 0.1 to 3.2 ± 0.4 in females (all p < 0.05 versus placebo).
  • In gonadectomized mice, β-ecdysone partially prevented trabecular bone loss; ORX + β-ecdysone had BV/TV 0.12 ± 0.01 versus 0.09 ± 0.01 in ORX, and OVX + β-ecdysone increased BFR/BS to 0.82 ± 0.2 versus 0.66 ± 0.2 in OVX (p < 0.05) and decreased CTX-1 to 20 ± 2.3 versus 24 ± 1.6 in OVX (p < 0.05), but did not benefit cortical bone in hormone-deficient mice.

Methods

  • Randomized animal study: 2-month-old male and female Swiss-Webster mice, 8 to 11 per group, received vehicle or β-ecdysone 0.5 mg/kg for 3 weeks; a separate experiment used gonadectomized male (ORX) and female (OVX) mice with sham-operated controls and β-ecdysone treatment.
  • Outcomes included micro-CT of the fifth lumbar vertebra and right midfemur, bone histomorphometry with calcein labeling, serum bone turnover markers (osteocalcin, CTX-1, osteoprotegerin, Dkk1), and biomechanical testing (lumbar vertebral compression and femoral three-point bending).
  • Analysis used two-way ANOVA with Bonferroni post hoc tests; p < 0.05 was significant, and sample size was based on prior glucocorticoid-induced bone loss data (eight animals per group for 80% power).

Limitations

  • Stated: only one β-ecdysone dose and one time point were tested; the study did not evaluate activation/deactivation or sensitivity of estrogen and androgen receptors; β-ecdysone failed to benefit cortical bone in hormone-deficient mice; longer or dose-dependent studies are needed.
  • Evident: this is a short-term mouse study, so human efficacy is not established; some comparisons lack reported significance (e.g., female vertebral maximum load); cortical bone histomorphometry was not performed for ORX/OVX + β-ecdysone groups because micro-CT and mechanical testing showed no structural or strength changes.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1007/s11999-015-4246-5

Cites

47 of 47 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 19 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Beta-ecdysone treatment did not alter body mass changes in male or female mice.

    “bEcd treatment did not alter body mass changes in the male or female mice (Table 1).”
    Quote in the paperNot a substantial claim
  • Findingown data

    In female mice, MS/BS did not change among the groups.

    “In female mice, MS/BS did not change among the groups.”
    Quote in the paperNot a substantial claim
  • Findingown data

    In male mice, osteoclast surface did not change among the treatment groups.

    “The osteoclast surface did not change among the treatment groups in male mice.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Female mice had similar vertebral bone mass and strength as male mice at 3 months of age.

    “We found that female mice had similar vertebral bone mass and strength as the male mice at 3 months of age.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Beta-ecdysone treatment at the current dose and treatment length failed to have beneficial effects on cortical bone in hormone-deficient mice.

    “However, bEcd treatment at the current dose and treatment length failed to have beneficial effects on the cortical bone in hormone-deficient mice.”
    Quote in the paperSubstantial claim
  • The quote only partly states this claim. 5
  • May combine two claims. 15
  • Classified as probably not a substantial claim. 16

  • The quote only partly states this claim.

    Findingown data

    In male mice, the orchiectomy plus beta-ecdysone group had higher trabecular bone volume than the orchiectomy alone group (0.12 ± 0.01 vs 0.09 ± 0.01).

    “and 0.12 ± 0.01 in the ORX + bEcd group.”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    In female mice, vertebral maximum load was 45 ± 2 in the placebo group and 48 ± 4 in the beta-ecdysone-treated group (not significant).

    “Maximum load of the vertebrae was 45 ± 2 in PL and 48 ± 4 in the bEcd-treated group”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The mice used gained approximately 10% body mass during the study period.

    “The mice we used gained approximately 10% body mass during the study period.”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    In male mice, the orchiectomy plus beta-ecdysone group had higher vertebral maximum load than the orchiectomy alone group (37 ± 3 vs 30 ± 4, p < 0.05).

    “and 37 ± 3 in the ORX + bEcd group.”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    In female mice, the ovariectomy plus beta-ecdysone group had trabecular bone volume of 0.14 ± 0.01, not higher than the ovariectomy alone group (0.15 ± 0.01).

    “and 0.14 ± 0.01 in the OVX + bEcd group.”
    Quote in the paperNot a substantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    In female mice, the ovariectomy plus beta-ecdysone group had vertebral maximum load of 44 ± 4, not significantly higher than the ovariectomy alone group (39 ± 4).

    “and 44 ± 4 in the OVX + bEcd group.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Sex differences in peak bone mass are largely predisposed by genetic determinants in rodents and humans.

    “Although sex differences in PBM are largely predisposed by genetic determinants in rodents [31] and humans [32], increased physical activities during the prepubertal period may alter PBM.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [31]

      Not checked in the free run.

    • cites [32]

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Epidemiological studies have shown that males have higher peak bone mass than females, which may delay osteoporosis development in males until their eighth decade of life.

    “Epidemiological studies have shown that males have higher PBM than females, which may delay the osteoporosis development in males until their eighth decade of life [2,5,16].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [2]

      Not checked in the free run.

    • cites [5] source

      Not checked in the free run.

    • cites [16]

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Two-month-old male and female Swiss-Webster mice were randomized to receive either vehicle or beta-ecdysone (0.5 mg/kg) for 3 weeks.

    “Two-month-old male and female Swiss-Webster mice were randomized to receive either vehicle or bEcd (0.5 mg/kg) for 3 weeks.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Beta-ecdysone is produced by arthropods to initiate metamorphosis, the so-called molting process.

    “One of these naturally derived products is b-Ecdysone (bEcd), which is produced by arthropods to initiate metamorphosis, the so-called molting process [36].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [36]

      Not checked in the free run.

  • Classified as probably not a substantial claim. May combine two claims.

    Study design and conductown data

    Sample size calculation was estimated from published data on glucocorticoid-induced bone loss using trabecular bone volume as the primary endpoint; the effective sample size to have 80% power and alpha 0.05 was eight animals per group.

    “Sample size calculation was estimated from our published data on glucocorticoid-induced (GC) bone loss using trabecular bone volume as our primary endpoint.Mean trabecular bone volume for the placebo-treated (PL) group was 12%, in GC was 10%, and sigma was 2%.The effective sample size to have 80% power and alpha 0.05 was eight animals per group.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductnone given

    All animals were treated according to the USDA animal care guidelines with the approval of the UC Davis Committee on Animal Research.

    “All animals were treated according to the USDA animal care guidelines with the approval of the UC Davis Committee on Animal Research.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Estrogen has effects on bone cell activity including periosteal apposition inhibition, endosteal bone resorption, and chondrocyte remodeling, resulting in epiphyseal closure and attenuated periosteal expansion.

    “Estrogen has a number of effects on bone cell activity including periosteal apposition inhibition, endosteal bone resorption, and chondrocyte remodeling.These effects result in epiphyseal closure and attenuate periosteal expansion [10].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10]

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    During puberty in males, serum levels of estrogen also increase, resulting in periosteal expansion through estrogen receptor alpha, whereas androgen acts on osteoblasts and osteocytes through androgen receptor to increase trabecular bone formation.

    “During puberty in males, serum levels of estrogen also increase, resulting in periosteal expansion through estrogen receptor a, whereas androgen acts on osteoblasts and osteocytes through androgen receptor to increase trabecular bone formation [27].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [27]

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    The beta-ecdysone dose was chosen from a pilot dose-response study performed on male mice with doses of 0.25 to 1 mg/kg.

    “The b-Ecd dose was chosen from a pilot dose-response study we performed on male mice with doses of 0.25 to 1 mg/kg [12].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [12]

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Achyranthes extracts are highly rich in beta-ecdysone.

    “Achyranthes extracts are highly rich in bEcd [7].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [7]

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    The study did not evaluate the potential activation or deactivation of hormonal receptors that include estrogen and androgen receptors after beta-ecdysone treatment or if beta-ecdysone would alter the sensitivities of the hormonal receptors.

    “Additionally, we did not evaluate the potential activation or deactivation of hormonal receptors that include estrogen and androgen receptors after bEcd treatment or if b-Ecd would alter the sensitivities of the hormonal receptors.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The bone development in mice in this study might resemble late pubertal human bone development.

    “For this reason, the bone development in mice in this study might resemble late pubertal human bone development.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Beta-ecdysone may differ from other anabolic hormone treatments that have caused serious cardiac complications when taken from the laboratory to human models.

    “bEcd may differ from the other anabolic hormone treatments that have caused some real problems when taken from the laboratory to human models such as serious cardiac complications.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data

    The study only used one dose of beta-ecdysone and only included one time point.

    “First, we only used one dose of b-ecdysone and only included one time point.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Although females had higher trabecular bone volume within the vertebral body, the smaller bone size in female mice compared with male mice may correspond to a lower maximum compression load and overall bone strength for both trabecular bone and cortical bone.

    “Although the females had higher trabecular bone volume within the vertebral body, the smaller bone size in female mice compared with the male mice may correspond to a lower maximum compression load and the overall bone strength (work to failure) for both trabecular bone (vertebrae) and cortical bone (femoral shafts).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Short-term administration of beta-ecdysone in growing intact mice resulted in marked enhancements in both trabecular and cortical bone formation that was associated with significantly increased trabecular and cortical bone volume.

    “In summary, we found that the short-term administration of bEcd in growing intact mice resulted in marked enhancements in both trabecular and cortical bone formation that was associated with significantly increased trabecular and cortical bone volume.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Beta-ecdysone did not bind to the estrogen receptor, and its pharmacological effects may not involve estrogen receptor activation.

    “However, it was reported that bEcd did not bind to the estrogen receptor [35], and its pharmacological effects may not involve estrogen receptor activation.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35]

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Treatment with beta-ecdysone increased both trabecular and cortical bone volume and bone strength in both sexes through anabolic mechanisms in gonadal-sufficient mice.

    “Treatment with bEcd increased both trabecular and cortical bone volume and bone strength in both sexes through anabolic mechanisms in gonadal-sufficient mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Beta-ecdysone may partially prevent the detrimental effects of gonadectomy on trabecular bone but not on cortical bone.

    “Our results indicate that bEcd may partially prevent the detrimental effects of gonadectomy on trabecular bone but not on cortical bone.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Beta-ecdysone treatment partially prevented trabecular bone loss primarily through further increases in tissue-level bone formation markers and bone formation rates.

    “bEcd treatment partially prevented trabecular bone loss primarily through further increases in tissue-level bone formation markers and bone formation rates.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Bone histomorphometry was not performed for the cortical bone from the orchiectomy and ovariectomy plus beta-ecdysone treatment groups as a result of the lack of structural and bone strength changes measured by micro-CT and mechanical testing.

    “Bone histomorphometry was not performed for the cortical bone from the ORX and OVX + bEcd treatment groups as a result of the lack of structural and bone strength changes measured by micro-CT and mechanical testing.”
    Quote in the paperNot a substantial claim
27 of 27

  • The paper relies on this without support.

    One of the strongest predictors for osteoporosis is peak bone mass.

    “One of the strongest predictors for osteoporosis is peak bone mass.”

  • The paper relies on this without support.

    Interventions to augment peak bone mass have yet to be developed.

    “Interventions to augment peak bone mass have yet to be developed.”

  • The paper relies on this without support.

    Beta-ecdysone has androgenic effects.

    “b-Ecdysone (bEcd), a natural steroid-like compound produced by arthropods to initiate metamorphosis, is believed to have androgenic effects and so may be used to augment bone mass.”
3 of 3

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • 20E alone increased bone formation, and when added concurrently with glucocorticoid, it compensated for the detrimental effect and prevented GC-induced autophagy in bone marrow stromal cells.

    “Moreover, 20E alone increased bone formation, and when added concurrently with GC, it compensated for the detrimental effect and prevented the GC-induced autophagy in bone marrow stromal cells, making it a promising neutralizer of GC side effects.”

    From Molecular Targets of 20-Hydroxyecdysone in Mammals, Mechanism of Action: Is It a Calorie Restriction Mimetic and Anti-Aging Compound? (2025) · cites this paper as [33]

    The Results section, “Effects of bEcd on Cortical Bone Formation in Gonadal-sufficient Mice” (page not provided), reports increased bone formation with β-ecdysone alone. The paper does not report concurrent glucocorticoid treatment or prevention of glucocorticoid-induced autophagy in bone marrow stromal cells; its glucocorticoid-related study appears only in the References and as background for the sample-size calculation. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • The beneficial effect of 20E alone on bone formation was observed in both sexes.

    “The beneficial effect of 20E alone on bone formation was observed in both sexes [33].”

    From Molecular Targets of 20-Hydroxyecdysone in Mammals, Mechanism of Action: Is It a Calorie Restriction Mimetic and Anti-Aging Compound? (2025) · cites this paper as [33]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • OVX trabecular bone loss was not prevented

    Major

    The discussion says bEcd partially prevented trabecular bone loss in OVX mice, but the reported trabecular bone volume was lower with bEcd than with vehicle (0.14 versus 0.15).

  • Female vertebral result is assigned to an ORX group

    Minor

    The female vertebral work-to-failure result names an ORX group, although the paper defines ORX as the male gonadectomy and OVX as the female gonadectomy.

2 of 2

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Authors
Dai, W., Zhang, H., Zhong, Z. A., Jiang, L., Chen, H., Lay, Y.-A. E., Kot, A., Ritchie, R. O., Lane, N. E., & Yao, W.
Affiliations
  • Longhua Hospital Shanghai University of Traditional Chinese Medicine (CN)
  • Central South University (CN)
  • University of California Davis Medical Center (US)
  • Dalian Medical University (CN)
  • University of California, Berkeley (US)
ORCID
Venue
Clinical Orthopaedics & Related Research
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2015-08
Volume
473
Issue
8
Pages
2495-2504
ISSN
0009-921X (ISSN-L)
DOI
10.1007/s11999-015-4246-5
PMC
PMC4488193
OpenAlex
W2151373614
Cited by
15
References
47
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2022: 2 · 2021: 3 · 2020: 3 · 2019: 1 · 2017: 2
FWCI
0.69
Type
journal article
Language
en
Open access
Locations
  • Clinical Orthopaedics and Related Research (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • Natural Science Foundation of Shanghai
  • National Natural Science Foundation of China
  • Shanghai University of Traditional Chinese Medicine
  • Longhua Hospital Shanghai University of Traditional Chinese Medicine
  • National Institutes of Health
  • University of California, Davis
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (P50AR063043, K24AR048841, R01AR061366)
Topics
  • Health Sciences › Medicine › Orthopedics and Sports Medicine › Bone health and osteoporosis research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
Keywords
20-hydroxyecdysone, peak bone mass, trabecular bone volume, bone strength, sex differences, gonadectomy, orchiectomy, ovariectomy, osteoporosis, androgenic effects, skeletal development, vertebral bone, bone mineral density, bone microarchitecture, mice, bone resorption, osteoblasts, osteoclasts
MeSH terms
Animals, Biomechanical Phenomena, Ecdysterone, Female, Femur, Lumbar Vertebrae, Male, Orchiectomy, Osteogenesis, Ovariectomy, Sex Factors, Bone Density, Bone Remodeling, Mice, X-Ray Microtomography

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