Meng-Meng Chen

Effects of Growth Hormone and Testosterone on Cortical Bone Formation and Bone Density in Aged Orchiectomized Rats

Osteoporosis in men is a disease that is increasing in incidence, and with an increasing elderly population it poses a serious health problem. Since both testosterone (T) and growth hormone (GH) have an anabolic effect on bone and both decrease with aging, we were prompted to test whether the administration of these hormones in combination would increase bone mass in orchiectomized (orx) senile rats more than administration of either agent alone. Twenty-month-old male Wistar rats were divided into five groups with seven animals each: (a) age-matched intact control, (b) orx, (c) orx+GH (2.5 mg/kg/day), (d) orx+T [10 mg/kg, subcutaneous (s.c.), injection given twice a week], and (e) orx+GH+T. Testosterone and GH were given subcutaneously for 4 weeks. Bone histomorphometry of the tibial shaft showed that the orx group had lower cortical bone area than the intact control group. The decrease in cortical bone area was due to increased intracortical porosis as well as decreased periosteal bone formation rate (BFR). Administration of T to the orx animals prevented the development of the porosis and the decrease in periosteal BFR. The bone mineral content (BMC) and bone mineral density (BMD) of the femur as tested by dual-energy X-ray absorptiometry were significantly higher in the orx+T than in the orx group and were not significantly different from that of the intact control group. Administration of GH to the orx rats increased periosteal BFR significantly; however, the BMC and BMD measured were not increased significantly in comparison to the orx group. When GH and T were combined in treatment, the cortical bone area, periosteal BFR, and femoral BMD were all significantly higher than that of the orx and even higher than the intact control rats. Two-way analysis of variance shows that the individual effect of GH and T treatment on the periosteal BFR and cortical bone area was significant. The effect of T, but not GH, on femoral BMC and BMD was also significant; however, there is no synergistic interaction between the two treatments. Four weeks of orx with or without GH or T administration had no significant effect on tibial metaphyseal cancellous bone volume. In conclusion, this short-term study suggests that the combined intervention of GH and T in androgen-deficient aged male rats may have an independent effect in preventing osteopenia. The significant effect of GH+T may be attributed to the prevention of intracortical porosis, and an increase in periosteal bone formation and cortical bone mass.

Effect of Treadmill Exercise on Tibial Cortical Bone in Aged Female Rats: A Histomorphometry and Dual . Energy X-Ray Absorptiometry Study

The aim of this study was to increase our understanding of the effect of exercise on cortical bone mass and turnover in aged female rats. Female Sprague-Dawley rats, 14 months of age, were divided into four groups: 8 controls and 10 exercised for the 9-week study, and 8 controls and 9 exercised for the 16-week study. Exercise consisted of treadmill running at 17 m/min for one h/day and 5 days/week for 9 and 16 weeks. All animals received double fluorochrome labeling of bone prior to sacrifice. Histomorphometric analysis was performed on 30-microns-thick Villanueva-stained, undecalcified cross-sections of the tibial shaft. Tibial diaphyseal mineral density of each rat in the 16-week study was measured by dual energy x-ray absorptiometry in vivo at 0, 9, and 16 weeks. The diaphyseal mineral density of the exercised group was significantly greater than that of the control group (p < 0.05 by two-way ANOVA) and the individual slopes of the density vs. time was found to be higher in the exercised than in the control animals (mean +/- SE of exercised 0.56 +/- 0.13 vs. control 0.19 +/- 0.07 mg/cm2/week, p < 0.05) by the end of the experiment. The results of the histomorphometric analysis after 9 weeks of exercise showed that the periosteal labeled surface, mineral apposition rate, and bone formation rate were profoundly increased by 192% (p < 0.001), 35%, and 206% (p < 0.01), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth hormone administration potentiates the effect of treadmill exercise on long bone formation but not on the vertebrae in middle-aged rats

To determine whether growth hormone administration would potentiate bone response to the stimulation of exercise, 80 female rats aged 14 months were divided into control (CON), ovine growth hormone administration (0.5 mg/kg daily) (GH), treadmill exercise (17 m/minute, 60 minutes daily) (EX), and GH+EX groups for 9 and 16 weeks. Static and dynamic histomorphometry were measured on the tibial shaft and (L-5) vertebral cortical bone. The periosteal and endocortical bone formation rate of the tibial shaft were higher in both EX and GH+EX than in the CON group in the 9-week study. There is a synergistic interaction between the two interventions in both cortical surfaces. After 16 weeks of study, the cortical bone area and periosteal bone formation rate were higher only in the EX than in the CON group. In the L-5 vertebra, the labeled surface on the periosteum was higher in the EX and the bone formation rate on the endocortical surface was higher in the GH than in the CON group. However, there was a negative interaction when the two interventions were combined. We conclude that a low-dose of growth hormone administration could initially potentiate long bone response to exercise. However, from the present study, long-term treatment with low-dose growth hormone administration does not enhance the increase in bone mass from exercise.

Ovariectomy-induced high turnover in cortical bone is dependent on pituitary hormone in rats

The aim of this study is to examine the interrelationship of pituitary and ovarian hormone deficiency on the regulation of bone growth and bone formation rate. 48 female rats, at 3 months of age, were divided into age-matched intact control, hypophysectomized (HX), ovariectomized (OV), and HX + OV groups. Ten rats were killed at 3 months of age as baseline controls, and the rest of the animals were killed 5 weeks after surgery. Serum levels of osteocalcin and dynamic histomorphometry on the periosteal surface of the tibial shaft and fifth lumbar vertebrae were measured to evaluate systemic and local bone turnover. Tibial and fourth lumbar vertebral bone area, bone mineral content, and bone density were measured by dual-energy X-ray absorptiometry (DXA). Our results confirmed that OV increased and HX suppressed systemic and periosteal bone formation parameters in both bone sites, OV increased and HX suppressed the gain in bone size and bone mass. When OV rats were HX, the serum levels of osteocalcin and periosteal bone formation parameters of the tibial shaft and the fifth lumbar vertebrae were, however, depressed and did not differ from that of the HX alone. DXA results show that the effect of OV on bone size and bone mass is also abolished by HX. In conclusion, we have demonstrated that OV increases tibial and lumbar vertebral bone formation and bone growth and this effect is pituitary hormone dependent.

Effects of estrogen and growth hormone on skeleton in the ovariectomized rat with hypophysectomy.

To investigate whether growth hormone (GH) and 17beta-estradiol (E2) replacement can prevent osteopenia induced by pituitary and ovarian hormone deficiency [by hypophysectomy and ovariectomy (HX+OV)], we administered relatively low doses of GH (2.3 IU x kg(-1) x day(-1)) and E2 (100 microg x kg(-1) x wk(-1)) in experiment 1 and relatively high doses of GH (13.5 IU x kg(-1) x day(-1)) and E2 (3,500 microg x kg(-1) x wk(-1)) in experiment 2 to 2-mo-old HX+OV Sprague-Dawley rats for 6 wk. Our data show that the HX+OV of rats results in diminished periosteal bone formation, longitudinal bone growth, and decreased cancellous bone volume. Administration of either the low or high dose of GH to these rats increased their systemic growth, serum levels of osteocalcin, and cortical bone formation. Either low or high doses of GH or E2 alone only partially prevent cancellous bone loss. However, the combined treatment of GH plus E2 resulted in an additive increase in the cancellous bone mass. We conclude that the additive effect of GH plus E2 on cancellous bone is attributed to the suppressive effect of E2 on bone resorption and the anabolic effect of GH on bone formation.To investigate whether growth hormone (GH) and 17β-estradiol (E2) replacement can prevent osteopenia induced by pituitary and ovarian hormone deficiency [by hypophysectomy and ovariectomy (HX+OV)], we administered relatively low doses of GH (2.3 IU ⋅ kg-1 ⋅ day-1) and E2 (100 μg ⋅ kg-1 ⋅ wk-1) in experiment 1 and relatively high doses of GH (13.5 IU ⋅ kg-1 ⋅ day-1) and E2 (3,500 μg ⋅ kg-1 ⋅ wk-1) in experiment 2 to 2-mo-old HX+OV Sprague-Dawley rats for 6 wk. Our data show that the HX+OV of rats results in diminished periosteal bone formation, longitudinal bone growth, and decreased cancellous bone volume. Administration of either the low or high dose of GH to these rats increased their systemic growth, serum levels of osteocalcin, and cortical bone formation. Either low or high doses of GH or E2 alone only partially prevent cancellous bone loss. However, the combined treatment of GH plus E2 resulted in an additive increase in the cancellous bone mass. We conclude that the additive effect of GH plus E2 on cancellous bone is attributed to the suppressive effect of E2 on bone resorption and the anabolic effect of GH on bone formation.

Effects of 17β-estradiol administration on cortical and cancellous bone of ovariectomized rats with and without hypophysectomy

Pituitary hormones are essential for bone growth and bone turnover. Hypophysectomy (HX) diminishes mitogenesis and abolishes the high bone turnover rate induced by ovariectomy (OV). It is not known whether the suppressive effect of estrogen on bone resorption is diminished or abolished by HX. The present study investigates the effects of 17 beta-estradiol (E2) (20 micrograms/wk) on cortical and cancellous bone mass and bone turnover as measured by histomorphometry in HX + OV (HO) rats. Sprague-Dawley rats at 2 months of age were OV or HO and the experiment was performed over a 6 week period. Hypophysectomy + OV (HO) resulted in a cessation of periosteal bone formation, and longitudinal bone growth and a decrease in cancellous bone volume. The tibial dry weight and tibial density were significantly lower in the HO than in the intact or OV groups. Administration of E2 to HO rats partially prevented cancellous bone loss, whereas the same dosage of E2 fully prevented cancellous bone loss in rats with OV alone. Nevertheless, cancellous bone volume was higher in the HO + E2 than in the HO-alone groups. Estradiol administration in HO rats did not suppress cancellous bone formation rate or the eroded surface as much as it did in the OV rats. The suppressive effect of E2 on periosteal bone formation rate and mineral apposition rate was also diminished in HO rats. However, factorial ANOVA showed that the effects of E2 on increasing cancellous bone volume and decreasing periosteal bone formation rate and mineral apposition rate were still significant in the HO rats. Tibial dry weight and tibial density did not differ between HO and HO + E2 groups. In conclusion, we have demonstrated that the estrogen-induced effects of preventing cancellous bone loss, of suppressing bone formation, and resorption as seen in OV rats was diminished but not abolished in HO rats.

Histologic evidence: Growth hormone completely prevents reduction in cortical bone gain and partially prevents cancellous osteopenia in the tibia of hypophysectomized rats

In previous studies we found that the cause of bone loss in young hypophysectomized (HX) animals was due primarily to an inhibition in growth‐dependent bone gain and a decrease in bone turnover. The aim of this study was to determine whether growth hormone, which has stimulatory effects on bone growth and turnover, can prevent HX‐induced skeletal alterations in rats.

Effect of hypophysectomy on the proliferation and differentiation of rat bone marrow stromal cells

Conditions such as estrogen deficiency, skeletal unloading, and aging have all been demonstrated to have various effects on the proliferation and differentiation of bone marrow stroma-derived osteoprogenitor cells. Here we have sought to examine the effects of pituitary hormone deficiency on the proliferation and the differentiation of these osteoprogenitor cells using the hypophysectomized (HX) rat as a model. In the present study, we use an in vitro culture system to examine the effects of HX on the osteogenic potential of rat bone marrow stroma. With the intact animal as a control, we used [3H]thymidine incorporation and cell number as indexes of proliferation. We also measured alkaline phosphatase enzyme activity, relative levels of osteocalcin expression with RT-PCR, and osteopontin and bone sialoprotein steady-state levels by Northern blot to delineate the effect on differentiation. Our results indicate that osteoprogenitor cells exposed to a pituitary hormone-deficient environment in vivo demonstrate an enhanced proliferative capacity and also exhibit an augmented expression of differentiation markers when exposed to an optimal environment in vitro.Conditions such as estrogen deficiency, skeletal unloading, and aging have all been demonstrated to have various effects on the proliferation and differentiation of bone marrow stroma-derived osteoprogenitor cells. Here we have sought to examine the effects of pituitary hormone deficiency on the proliferation and the differentiation of these osteoprogenitor cells using the hypophysectomized (HX) rat as a model. In the present study, we use an in vitro culture system to examine the effects of HX on the osteogenic potential of rat bone marrow stroma. With the intact animal as a control, we used [3H]thymidine incorporation and cell number as indexes of proliferation. We also measured alkaline phosphatase enzyme activity, relative levels of osteocalcin expression with RT-PCR, and osteopontin and bone sialoprotein steady-state levels by Northern blot to delineate the effect on differentiation. Our results indicate that osteoprogenitor cells exposed to a pituitary hormone-deficient environment in vivo demonstrate an enhanced proliferative capacity and also exhibit an augmented expression of differentiation markers when exposed to an optimal environment in vitro.

Anabolic Effect of Prostaglandin E2 in Bone Is not Dependent on Pituitary Hormones in Rats

Prostaglandin E2 (PGE2) is an anabolic agent of bone in vivo but the mechanism of its action still remains unclear. The aim of this study was to determine whether the effect of PGE2 on skeleton is mediated by pituitary hormones. Forty female, Sprague-Dawley rats were divided into four groups: baseline control (basal), age-matched intact control (CON), hypophysectomy (HX), and HX + PGE2 (2 mg/kg/day) with 10 animals in each group. The basal group was sacrified at 2 months of age, and the remaining groups after 6 weeks of treatment. Cancellous and cortical bone histomorphometry was performed on double fluorescent-labeled 40 μm-thick sections of the proximal tibia and tibial shaft. Our results show that HX resulted in a cessation of bone growth, a decrease in cancellous bone volume, and cortical bone gain compared with the age-matched, intact CON rats. Compared with the HX group, the HX + PGE2 group had a significantly greater tibial bone density (mean ± SE, HX + PGE2:1.595 ± 0.007 versus HX:1.545 ± 0.013), percent cancellous bone volume (21.4 ± 2.0 versus 8.41 ± 1.70), percent cortical bone area (87.2 ± 0.85 versus 81.7 ± 0.7), and ratio of cortical area to marrow area (7.14 ± 0.56 versus 4.52 ± 0.21). Increased bone masses by PGE2 in the HX animals were accompanied by an increase in the trabecular and endosteal-labeled surface and bone formation rate. The trabecular number and width were increased whereas trabecular separation was decreased in the HX + PGE2 group compared with the HX group (P < 0.05). PGE2 treatment also caused a decrease in the tibial endosteal eroded surface and medullar cavity of the HX animals. In conclusion, this study clearly demonstrates that PGE2 (2 mg/kg/day) in the HX rats increases both cortical and cancellous bones and improves trabecular architecture in the tibia after 6 weeks of treatment. These skeletal alterations are due to a stimulation of bone formation and a suppression of bone resorption activity. These findings suggest that the anabolic effect of PGE2 in bone is independent of pituitary hormones.