The effect of maternal HMB supplementation on bone mechanical and geometrical properties, as well as histomorphometry and immunolocalization of VEGF, TIMP2, MMP13, BMP2 in the bone and cartilage tissue of the humerus of their newborn piglets

Abstract

The presented experiment focuses on assessing the impact of HMB (hydroxy-β-methobutyrate) supplementation of mothers during pregnancy on the development of the skeletal system of their offspring. For this purpose, an experiment was carried out on 12 clinically healthy sows of the Great White Poland breed, which were divided randomly into two groups the control and the HMB group. All animals were kept under standard conditions and received the same feed for pregnant females. In contrast, females from the HMB group between 70 and 90 days were supplemented with 3-hydroxy-3-methylbutyle in the amount of 0.2g/kg b.w/day. Immediately after birth, the piglets were also divided into groups based on: sex, and presence or lack HMB supplementation, and subsequently were euthanized and humerus bones from all piglets were collected. Mother’s HMB supplementation during pregnancy affected the multiple index of their offspring. The higher humerus mass and length was observed with the greater effect in males. Maternal supplementation also influenced on the geometrical and mechanical properties of the humerus as in the case of mass, this effect was higher in males. Also, the collagen structure of the compacted and trabecular bone changed under the HMB addition. Maternal supplementation also affected the expression of selected proteins in growth cartilage and trabecular bone. The obtained results show that the administration to the mother during pregnancy by the HMB significantly affects the development of the humerus in many ways. The obtained results also confirm the utility of such experiments in understanding of the importance of the pregnancy diet as an develop and adaptable factor of offspring organisms and are the base for further research in that area as well as in the protein markers expression area.

Femoral µCT Analysis, Mechanical Testing and Immunolocalization of Bone Proteins in β-Hydroxy β-Methylbutyrate (HMB) Supplemented Spiny Mouse in a Model of Pregnancy and Lactation-Associated Osteoporosis

“…Negative control sections for each antibody were conducted by identical immunohistochemical staining excluding the primary antibody ( Supplementary Data, Figure S1 ). The intensity of immunoreaction was measured using a CX43 microscope (Olympus, Tokyo, Japan), both by determining the percentage of cells with a positive response, and by the quantitative comparison of mean pixel intensity in the photomicrographs, which were firstly converted into negatives and then into 8-bit grey-scale digital images, with a scale from 0 (white pixel) to 255 (black pixel), where the higher the pixel value, the higher the intensity of the immunohistochemical reaction [ 29 , 44 ]. For the immunoreactive cell count procedure, four randomly selected areas of compact bone, trabecular bone and articular cartilage, respectively, were measured for each microscopic slide.…”

“…Functional food or supplements affecting skeletal tissue remodeling and homeostasis can be useful for prevention before PLO. Based on previous results, HMB supplementation during pregnancy has been shown to have a positive effect, not only on the offspring, but also on the pregnant dam’s skeletal system [ 21 , 27 , 28 , 29 ]. Taking into account the influence of pregnancy on bone quality and the effect of HMB in protecting pregnant women against excessive bone loss, it seems reasonable to investigate the effects of HMB supplementation during pregnancy on the immunolocalization of selected proteins involved in bone metabolism in pregnancy and lactation.…”

Yolk Fatty Acid Profile and Amino Acid Composition in Eggs from Hens Supplemented with ß-Hydroxy-ß-Methylbutyrate

“…Studies on pigs supplemented with HMB have shown that it regulates lipid metabolism, promoting fat oxidation, which could potentially improve body composition and aid in fat loss [22][23][24]. Additionally, HMB enhanced piglets bone health by promoting bone formation, reducing resorption, and increasing bone density [25][26][27][28].…”

Evaluation of the Multiple Tissue Factors in the Cartilage of Primary and Secondary Rhinoplasty in Cleft Lip and Palate Patients

“…Furthermore, irrespective of their MMP neutralizing actions, TIMPs are thought to have implications of pluripotency on cellular functions, such as cell proliferation, movement, endurance, and differentiation [ 13 ]. Tissue inhibitor of metalloproteinases 2 (TIMP-2) expression is lower in studies with pregnant pig mothers receiving hydroxy—methobutyrate supplementation for the skeletal development of their piglets, which could imply that these supporting tissues of animals are undergoing greater remodeling [ 14 ]. The effect of a combination of anabolic growth factors and a protease inhibitor on an in vitro culture of articular chondrocytes revealed that TIMP-2 was beneficial in enhancing ECM synthesis [ 15 ].…”