Hesperidin Promotes Osteogenesis and Modulates Collagen Matrix Organization and Mineralization In Vitro and In Vivo

Abstract

This study evaluated the direct effect of a phytochemical, hesperidin, on pre-osteoblast cell function as well as osteogenesis and collagen matrix quality, as there is little known about hesperidin’s influence in mineralized tissue formation and regeneration. Hesperidin was added to a culture of MC3T3-E1 cells at various concentrations. Cell proliferation, viability, osteogenic gene expression and deposited collagen matrix analyses were performed. Treatment with hesperidin showed significant upregulation of osteogenic markers, particularly with lower doses. Mature and compact collagen fibrils in hesperidin-treated cultures were observed by picrosirius red staining (PSR), although a thinner matrix layer was present for the higher dose of hesperidin compared to osteogenic media alone. Fourier-transform infrared spectroscopy indicated a better mineral-to-matrix ratio and matrix distribution in cultures exposed to hesperidin and confirmed less collagen deposited with the 100-µM dose of hesperidin. In vivo, hesperidin combined with a suboptimal dose of bone morphogenetic protein 2 (BMP2) (dose unable to promote healing of a rat mandible critical-sized bone defect) in a collagenous scaffold promoted a well-controlled (not ectopic) pattern of bone formation as compared to a large dose of BMP2 (previously defined as optimal in healing the critical-sized defect, although of ectopic nature). PSR staining of newly formed bone demonstrated that hesperidin can promote maturation of bone organic matrix. Our findings show, for the first time, that hesperidin has a modulatory role in mineralized tissue formation via not only osteoblast cell differentiation but also matrix organization and matrix-to-mineral ratio and could be a potential adjunct in regenerative bone therapies.

Systemic Dietary Hesperidin Modulation of Osteoclastogenesis, Bone Homeostasis and Periodontal Disease in Mice

“…Phytochemicals, particularly flavonoids, are gaining increased interest for their therapeutic potential for bone. Studies reporting the effects of flavonoids in cell culture and animal models have strongly supported a role for flavonoids in bone formation and resorption in vitro and in vivo [19,22,25,27]. Additionally, the intake of flavonoids has been shown to improve bone health not only due to an influence in osteogenesis and osteoclastogenesis, but also due to their antioxidant and anti-inflammatory properties [26,27].…”

“…In order to evaluate the effect of HE on osteoclasts and bone homeostasis in vivo, two studies were conducted: a model of inflammatory bone loss in the alveolar housing and a skeletal bone phenotype characterization after a long (determined as enough time to affect bone connective tissue and cell function-6 weeks and 12 weeks) dietary supplementation with HE. HE has been shown to affect bone mass in rodents primarily due to inhibition of osteoclastogenesis, although there are reports that it may increase the number of osteoblasts and increase osteoblast function at low doses as well [19,22,23,25,30]. The mouse model of periodontal inflammatory bone resorption used in our study was chosen based on an acute-to-chronic inflammatory response to interdentally placed ligature [42,43] without the introduction of virulent factors such as pathogenic periodontal bacteria.…”

“…It has also been suggested that HE has an additional protective effect on bone by modulating the production of inflammatory mediators [22]. Recently, our research evaluating the direct effect of HE on osteogenesis showed that HE has a positive role in mineralized tissue formation via not only promotion of osteoblast cell differentiation, but also improvement of matrix organization and matrix-to-mineral ratio, providing a basis to the idea that it could be a potential adjunct in regenerative and maintenance-driven bone therapies [25].…”

Hesperidin Ameliorates Dexamethasone-Induced Osteoporosis by Inhibiting p53

“…In a previous study, hesperidin protected bone mass loss by alleviating oxidative stress and inflammation in an ovariectomy rat model ( Zhang et al, 2021 ). Hesperidin could upregulate the expression of osteogenic markers and promote the maturation of bone organic matrix, thus exerting anti-osteoporosis effects in vitro and in vivo ( Miguez et al, 2021 ). In an experimental rat model, hesperidin intake resulted in bone mass gain in young rats and protected against ovariectomy-induced bone loss in adult rats, as well as reduced oxidative stress and total lipid content ( Horcajada et al, 2008 ).…”

Editorial: Tissue Stem Cells During Trauma: From Basic Biology to Translational Medicine

“…Hesperidin, a flavanone glycoside highly abundant in citrus fruits, has exhibited a protective role in osteoporosis through enhancing osteogenic differentiation and inhibition of osteoclastogenesis ( Liu et al, 2019 ; Miguez et al, 2021 ; Zhang et al, 2021 ), but little is known about the role of hesperidin in the dexamethasone-induced osteoporosis. Zhang et al indicated that hesperidin showed enhanced osteogenic differentiation and partially reversed dexamethasone-induced inhibition of osteogenic differentiation by p53 signaling pathway in vitro , suggesting the compound may be a promising candidate against dexamethasone-induced osteoporosis ( Zhang et al, 2021 ).…”