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Changes in the Fracture Resistance of Bone with the Progression of Type 2 Diabetes in the ZDSD Rat

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Abstract

Individuals with type 2 diabetes (T2D) have a higher fracture risk compared to non-diabetics, even though their areal bone mineral density is normal to high. Identifying the mechanisms whereby diabetes lowers fracture resistance requires well-characterized rodent models of diabetic bone disease. Toward that end, we hypothesized that bone toughness, more so than bone strength, decreases with the duration of diabetes in ZDSD rats. Bones were harvested from male CD(SD) control rats and male ZDSD rats at 16 weeks (before the onset of hyperglycemia), at 22 weeks (5–6 weeks of hyperglycemia), and at 29 weeks (12–13 weeks of hyperglycemia). There were at least 12 rats per strain per age group. At 16 weeks, there was no difference in either body weight or glucose levels between the two rat groups. Within 2 weeks of switching all rats to a diet with 48 % of kcal from fat, only the ZDSD rats developed hyperglycemia (>250 mg/dL). They also began to lose body weight at 21 weeks. CD(SD) rats remained normoglycemic (<110 mg/dL) on the high-fat diet and became obese (>600 g). From micro-computed tomography (μCT) analysis of a lumbar vertebra and distal femur, trabecular bone volume did not vary with age among the non-diabetic rats but was lower at 29 weeks than at 16 weeks or at 22 weeks for the diabetic rats. Consistent with that finding, μCT-derived intra-cortical porosity (femur diaphysis) was higher for ZDSD following ~12 weeks of hyperglycemia than for age-matched CD(SD) rats. Despite an age-related increase in mineralization in both rat strains (μCT and Raman spectroscopy), material strength of cortical bone (from three-point bending tests) increased with age only in the non-diabetic CD(SD) rats. Moreover, two other material properties, toughness (radius) and fracture toughness (femur), significantly decreased with the duration of T2D in ZDSD rats. This was accompanied by the increase in the levels of the pentosidine (femur). However, pentosidine was not significantly higher in diabetic than in non-diabetic bone at any time point. The ZDSD rat, which has normal leptin signaling and becomes diabetic after skeletal maturity, provides a pre-clinical model of diabetic bone disease, but a decrease in body weight during prolonged diabetes and certain strain-related differences before the onset of hyperglycemia should be taken into consideration when interpreting diabetes-related differences.

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Abbreviations

AGEs:

Advanced glycation end products

α-ABA:

Alpha-amino-N-butyric acid

aBMD:

Areal bone mineral density

Ct.Ar:

Bone cross-sectional area

BW:

Body weight

BV/TV:

Bone volume fraction

CD(SD):

CD IGS rat (Clr:CD(SD)) from Charles River

Conn.D:

Connectivity density

Ct.Th:

Cortical thickness

Ct.TMD:

Cortical tissue mineral density

K c,int :

Crack initiation toughness

DPD:

Deoxypyridinoline

DFM:

Distal femur metaphysis

C min :

Distance between centroid and periosteal surface

EDTA:

Ethylenediaminetetraacetic acid

HFBA:

Heptafluorobutyric acid

HFD:

High-fat diet

HPLC:

High-performance liquid chromatography

HR-pQCT:

High-resolution peripheral quantitative computed tomography

HA:

Hydroxyapatite

Ct.Po:

Intra-cortical porosity

μCT:

Micro-computed tomography

MMR:

Mineral-to-matrix ratio

I min :

Moment of inertia

NEGs:

Non-enzymatic, glycation mediated crosslinks

M p :

Peak moment

PE:

Pentosidine

PITC:

Phenylisothiocyanate

PBS:

Phosphate buffer saline

Po.N:

Pore number

PYdisp :

Post-yield displacement

PYD:

Pyridinoline

PYR:

Pyridoxine

RS:

Raman spectroscopy

ROI:

Region of interest

L:

Span

3pt:

Three-point bending

Tt.Ar:

Total cross-sectional area

Tb.N:

Trabecular number

Tb.Th:

Trabecular thickness

Tb.TMD:

Trabecular tissue mineral density

T2D:

Type 2 diabetes

VB:

Vertebral body

W f :

Work-to-fracture

ZDF:

Zucker diabetic fatty

ZDSD:

Zucker diabetic Sprague–Dawley from PreClinOmics

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Acknowledgments

The Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development (1I01BX001018) primarily funded the present work. The fund for the purchase of the micro-computed tomography scanner was provided by the National Center for Research Resources (1S10RR027631) and matching funds from the Vanderbilt Office of Research. There was additional support from the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health (NIH grants AR063157 and 1T32DK101003) and the National Science Foundation (1068988). The Vanderbilt Hormone Assay and Analytical Services Core is supported by NIH grants DK059637 and DK020593. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or other funding agencies.

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Authors and Affiliations

  1. Department of Veterans Affairs, Tennessee Valley Healthcare System, Nashville, TN, 37212, USA

    Amy Creecy, Sasidhar Uppuganti, Alyssa R. Merkel, Alexander J. Makowski, Mathilde Granke & Jeffry S. Nyman

  2. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37232, USA

    Amy Creecy, Alexander J. Makowski & Jeffry S. Nyman

  3. Center for Bone Biology, Vanderbilt University Medical Center, Nashville, TN, 37232, USA

    Amy Creecy, Sasidhar Uppuganti, Alyssa R. Merkel, Alexander J. Makowski, Mathilde Granke & Jeffry S. Nyman

  4. Department of Orthopaedic Surgery & Rehabilitation, Vanderbilt University Medical Center, 1215 21st Ave S., Suite 4200, Nashville, TN, 37232, USA

    Sasidhar Uppuganti, Mathilde Granke & Jeffry S. Nyman

  5. Department of Medicine, Division of Clinical Pharmacology, Vanderbilt University Medical Center, Nashville, TN, 37232, USA

    Alyssa R. Merkel

  6. School of Medicine, Meharry Medical College, Nashville, TN, 37208, USA

    Dianne O’Neal

  7. Department of Medicine, Division of Nephrology, Vanderbilt University Medical Center, Nashville, TN, 37232, USA

    Paul Voziyan

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  1. Amy Creecy
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Correspondence to Jeffry S. Nyman.

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Conflicts of interest

Amy Creecy, Sasidhar Uppuganti, Alyssa R. Merkel, Dianne O’Neal, Alexander J. Makowski, Mathilde Granke, Paul Voziyan, and Jeffry S. Nyman have declared no conflict of interest.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.

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Creecy, A., Uppuganti, S., Merkel, A.R. et al. Changes in the Fracture Resistance of Bone with the Progression of Type 2 Diabetes in the ZDSD Rat. Calcif Tissue Int 99, 289–301 (2016). https://doi.org/10.1007/s00223-016-0149-z

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