Abnormalities in Microarchitecture and Reduced Mechanical Bone Strength in a Rat Model of Polycystic Ovary Syndrome
Authors: Comim Fabio Vasconcellos , Mujica Lady Katerine Serrano , Valente Fernanda , Miyazato Ligia G , Manta Manuela W , Amaral Carolina D , Premaor Melissa O , Macari Soraia , Silva Tarcilia , Barrioni Breno R , Carlos Bruna L , Silva Guilherme J A , Shimano Antonio Carlos , Antoniazzi Alfredo Q 
Published: May 2021

Abstract
Evidence from the literature is contentious about the impact of polycystic ovary syndrome (PCOS) on the skeleton, suggesting a possible negative role of this condition on non-obese women. We investigated this hypothesis employing a well-characterized testosterone propionate (TP) rodent model of PCOS to address the consequences of androgenization on bone microarchitecture, histology, and mechanical strength. For this study, Wistar rats (n= 38) were divided in 4 groups: 1) “Control OVX” (single dose of corn oil s.c. at day 5 of life and ovariectomy at day 100, n=9); 2) “Control SHAM” (n=9); 3) “Androgenized OVX”(single dose of TP 1.25 mg s.c. at day 5 of life and ovariectomy at day 100, n=10); and 4) “Androgenized SHAM” (n=10). Full characterization of estrous cycles and weight was performed during growth, and all animals were euthanized at day 180. Successful ovariectomy was confirmed by neglected levels of serum estradiol. Endpoints evaluated include bone micro CT (femur and spinal column), bone histology (number of osteoclasts and osteoblasts in the femur), and mechanical tests. The study was approved by the local Ethics Committee. At the end of the study (day 180), Androgenized OVX rats were heavier than the other three groups. MicroCT Analysis: Androgenized SHAM rats exhibited a significantly higher trabecular mass in the spine (BV/BT) (mean + SEM) 49.21 + 2.42 % versus Control SHAM 36.42 + 1.39 % (Student T-test p=0.001). Following ovariectomy, BV/BT in Androgenized OVX was 40.4 + 2.83 % against 20.34 + 1.85 % in Control OVX (Student T-test p=0.0003). Lumbar trabecular thickness(μm) was also higher in Androgenized OVX (p=0.0065) as well the Trabecular number (n/mm)(p=0.0003). A similar increase in trabecular mass was observed in the femur. Androgenized SHAM rats had a significant higher BV/BT (%), trabecular thickness(μm), and decreased trabecular separation (p &lt; 0.001). However, a significant reduction in cortical bone (thickness) was noted (Student T-test p=0.001). A histological study of the distal femur of Androgenized SHAM rats also show a significantly increased number of osteoclasts and decreased number of osteoblasts than Control SHAM (0&lt; 001). When submitted to the mechanical test, Androgenized Sham rats presented a decreased strength (p&lt;0.01) in relation to its controls. After ovariectomy, there was a reduction in bone in all oophorectomized groups. However, differently than the vertebral bones, no differences regarding bone mechanical strength or stiffness as well microCT values, or bone histology parameters were noted in the femur of Control OVX or Androgenized OVX. Our results suggest that androgenization in a rodent model of PCOS leads, at the same time, to a generalized increase in trabecular (cancellous) bone mass (especially in the spine), associated with a reduced cortical bone mass and decreased strength of the femur.