Menaquinones, also known as the vitamin K2 family, regulate calcium homeostasis in a "bonevascular cross-talk" and have recently received particular attention for their positive effect on bone formation. Since the correlation between menaquinones and bone metabolism is not yet clear, we verified the effect of the MK-4 molecule, a type of menaquinone, in the modulation of osteogenesis. For this reason, we compared two-dimensional (2D) and three-dimensional (3D; RCCS-4™ bioreactor) in vitro culture models. In particular, to recreate the "bone remodeling unit" in vitro, human amniotic fluid mesenchymal stem cells (hAFMSC) were co-cultured in the 3D system with human monocytes (hMCs) cells as osteoclast precursors. The results showed that in a conventional 2D culture system, hAFMSCs responded to MK-4, which improved the osteogenic process through the γ-glutamyl carboxylase-dependent pathway. The same results were identified in the 3D dynamic system in which the MK-4 treatment supported osteoblast-like formation by promoting extracellular deposition of the bone matrix and the expression of osteogenic-related proteins (alkaline phosphatase, osteopontin, type 1 collagen and osteocalcin). In particular, when hAFMSCs were cultivated in co-culture in a 3D dynamic system with hMCs, the presence of MK-4 supported the formation of cellular aggregates and the osteogenic function of hAFMSC, but negatively influenced the process of osteoclastogenesis. Our results demonstrated that 3D hAFMSC/ hMC construct can be a useful cellular model for the in vitro screening of bioactive molecules in bone tissue. As we previously demonstrated(a) by using the same experimental culture model, 3D osteoblasts/osteoclasts co-culture can be generated employing limited amounts of human primary cells from the same patient, mimicking the bone microenvironment both in healthy or pathological condition. This will allow the establishment of a smart system providing information on the efficacy of pharmacological treatment aimed at considering the characteristics of each patient. Reference a. Penolazzi, L, Lolli, A, Sardelli, L, Angelozzi, M, et al.. Establishment of a 3D-dynamic osteoblasts-osteoclasts co-culture model to simulate the jawbone microenvironment in vitro. Life Sci. 2016 May 1;152:82-93.
Generation of 3D bone mimetic culture system for drug screening: the Vitamin K2 model.
Penolazzi L;Lambertini E;A Piva
2019
Abstract
Menaquinones, also known as the vitamin K2 family, regulate calcium homeostasis in a "bonevascular cross-talk" and have recently received particular attention for their positive effect on bone formation. Since the correlation between menaquinones and bone metabolism is not yet clear, we verified the effect of the MK-4 molecule, a type of menaquinone, in the modulation of osteogenesis. For this reason, we compared two-dimensional (2D) and three-dimensional (3D; RCCS-4™ bioreactor) in vitro culture models. In particular, to recreate the "bone remodeling unit" in vitro, human amniotic fluid mesenchymal stem cells (hAFMSC) were co-cultured in the 3D system with human monocytes (hMCs) cells as osteoclast precursors. The results showed that in a conventional 2D culture system, hAFMSCs responded to MK-4, which improved the osteogenic process through the γ-glutamyl carboxylase-dependent pathway. The same results were identified in the 3D dynamic system in which the MK-4 treatment supported osteoblast-like formation by promoting extracellular deposition of the bone matrix and the expression of osteogenic-related proteins (alkaline phosphatase, osteopontin, type 1 collagen and osteocalcin). In particular, when hAFMSCs were cultivated in co-culture in a 3D dynamic system with hMCs, the presence of MK-4 supported the formation of cellular aggregates and the osteogenic function of hAFMSC, but negatively influenced the process of osteoclastogenesis. Our results demonstrated that 3D hAFMSC/ hMC construct can be a useful cellular model for the in vitro screening of bioactive molecules in bone tissue. As we previously demonstrated(a) by using the same experimental culture model, 3D osteoblasts/osteoclasts co-culture can be generated employing limited amounts of human primary cells from the same patient, mimicking the bone microenvironment both in healthy or pathological condition. This will allow the establishment of a smart system providing information on the efficacy of pharmacological treatment aimed at considering the characteristics of each patient. Reference a. Penolazzi, L, Lolli, A, Sardelli, L, Angelozzi, M, et al.. Establishment of a 3D-dynamic osteoblasts-osteoclasts co-culture model to simulate the jawbone microenvironment in vitro. Life Sci. 2016 May 1;152:82-93.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
