The modified amino sugar N-Butyryl Glucosamine fed to ovariectomized rats preserves bone mineral through increased early mineralization, but does not affect body composition

Abstract

Background:  The toxicities of pharmaceuticals for chronic arthritis and osteoporosis should be of concern to consumers. This partially accounts for the popularity of consumption of the amino sugar glucosamine, in-spite of controversy about its efficacy. We chemically synthesized N-butyryl glucosamine (GlcNBu), which we discovered protected bone and cartilage in an inflammatory arthritis rat model when used as a feed supplement. GlcNBu can also be potentially synthesized biochemically, since we recently demonstrated that human acetyl-CoA: glucosamine-6-phosphate N-acetyltransferase 1 has a relaxed donor specificity and transfers acyl groups of up to four carbons in length, i.e. the butyryl moiety. Oral GlcNBu had no detectable toxicity and also protected against bone loss in ovariectomized (OVX) rats as a model for osteoporosis. However, we demonstrated this only for bones excised at 6 months. Thus, the current study aims to determine when bone mineralization is maximized during daily GlcNBu supplementation, in both OVΧ and Sham-OVX rats, in addition to the relationship of bone mineralization to body composition.

Methods: Female Sprague-Dawley rats were randomized into 4 groups, containing 8 rats each. The groups consisted of OVX or Sham-OVX rats whose diets were supplemented with either 200 mg/kg/day of GlcNBu or an equimolar amount of glucose. We performed sequential bone density and body composition measurements, by dual-energy X-ray absorptiometry in the live, anesthetised rats, over a 6-month experimental period, starting at the age of 8 weeks. Results were analyzed by descriptive statistics and 2-way ANOVA. 

Results: The major increases in the mineral content and density of the spine and the femur in GlcNBu-supplemented rats occurred early, from the baseline to week 8.  Ovariectomy resulted in a number of significant differences in body composition, while feeding GlcNBu had no significant effects on body composition. The significant effects of ovariectomy on body composition initially appeared at 8 weeks, while the GlcNBu effects on increased bone mineral initially appeared at 2 weeks. An interaction between OVX and GlcNBu was seen only at 16 weeks for the bone mineral density of the femoral head.

Conclusions: Supplementation of the diet by GlcNBu in both OVX and Sham-OVX rats increases bone mineral as early as 2 weeks. Ovariectomy but not GlcNBu supplementation had a significant effect on body composition. The effect of GlcNBu occurs independently of changes in body composition, probably as a direct effect of stimulation of bone matrix synthesis which continues to be mineralized.  This work represents an important step in the development and commercialization of GlcNBu for the prevention and treatment osteoporosis, where there is now an increasing demand for safe, long term agents.

Key words: osteoporosis, ovariectomy, N-butyryl glucosamine, bone, mineralization, body composition, dual-energy X-ray absorptiometry