Leukemia Inhibitory Factor and Oncostatin M Influence the Mineral Phases Formed in a Murine Heterotopic Calcification Model: A Fourier Transform-Infrared Microspectroscopic Study

Abstract

The study of bone mineralization processes is of considerable interest in understanding bone diseases and developing new therapies for skeletal disorders, particularly since bone homeostasis requires numerous cell types and a large cytokine network. Cell culture models of mineralization have often been used to study the cellular mechanisms of mineralization, but few data have been reported concerning the influence of extracellular matrix components and cytokines on the physicochemical properties of mineral. The purpose of this study was to analyze the effects of two cytokines, leukemia inhibitory factor (LIF) and oncostatin M (OSM), involved in bone metabolism on the physicochemical properties of bone mineral formed in a murine in vivo mineralization model. Murine bone marrow cells implanted under the kidney capsule in the presence or absence of cytokines led to heterotopic ossicle formation. A scanning electron microscopic microprobe revealed that heterotopic calcification had a lower (ϳ20%) Ca/P ratio after cytokine treatment as compared with the control without cytokine. Transmission electron microscopic analysis of cytokine-treated ossicles showed numerous areas with low mineral density, whereas electron diffraction pattern revealed an apatitic phase. These areas were not observed in the absence of cytokine. Moreover, Fourier transform-infrared microspectroscopy showed at the molecular level that the presence of either cytokine induced many microscopic areas in which short-range order organization, such as incorporation of carbonate and crystallinity/maturity of ossicle mineral, were modified. LIF and OSM influenced mineral phase formation in the present model and may thus be key protagonists in bone mineral development and skeletal diseases. (J Bone Miner Res 1998;13:1619-1632)

LIF Inhibits Osteoblast Differentiation at Least in Part by Regulation of HAS2 and Its Product Hyaluronan

“…(6) Finally, in a mouse model of heterotypic calcification, both LIF and OSM dose-dependently decreased the calcium/phosphate ratio of the mineral deposited on matrix formed by implanted murine bone marrow cells. (5) Taken together, these results suggest that LIF stimulates production of an abnormal osteoid that may contribute to altered recruitment and differentiation of osteoprogenitors, altered osteoblast activity, and abnormal mineralization.…”

“…Because the inhibitory effect of LIF on osteoprogenitor differentiation is irreversible and restricted to a relatively short sensitive window over the developmental time-course of RC cell cultures, (7,8) we designed a positive-negative differential display screening strategy to identify genes down-stream of LIF receptor signaling that may mediate the inhibition. To this end, RC cells were pulse-treated with either LIF or vehicle (PBS) for 3 consecutive days, within (days 7-10) and outside (days 10-13) the sensitive time window; as expected, LIF inhibited bone nodule formation only in cultures treated chronically (days [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] or in the inhibition-sensitive window, but not the insensitive window ( Fig. 1).…”

“…(1,2) Also, in a mouse model of heterotopic calcification, both human LIF and human oncostatin M (OSM), which can signal through the same heterodimeric receptor as LIF (gp130-gp190), (3,4) dosedependently decreased the calcium/phosphate ratio of the mineral deposited on matrix by implanted murine bone marrow cells. (5) Moreover, transgenic mice engineered to overexpress ubiquitously bovine OSM developed bone hypertrophy with immature osteoblasts, disrupted and disorganized growth plates, the formation of incomplete haversian systems, and no sign of remodeling. (6) Previously, we showed that LIF causes a dose-dependent decrease in the number of bone nodules formed in primary cultures of cells isolated from 21-day fetal rat calvaria (RC).…”

Bone regeneration: the stem/progenitor cells point of view

“…Within these conditions, cMSCs are defined as non‐haematopoietic cells (CD45 – , CD14 – , CD34 – ) expressing some molecules, the combination of which is largely used for their description: CD73 + , CD44 + , CD105 + , CD90 + and CD146 + (Table 1). Cultured MSCs are largely used in experimental bone reconstruction in vitro and in vivo [18, 23–25]. Indeed, several types of animal studies demonstrated their potential to produce bone both in the ectopic position and within the bone environment ( e.g.…”