Some factors affecting mineralization of bone in tissue culture

Ramp, Warren K.; Neuman, W. F.

doi:10.1152/ajplegacy.1971.220.1.270

Cited by 51 publications

(9 citation statements)

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“…An integral part of this hypothesis is the theory put forth by Ramp and Neuman [28] relating to the importance of an intact periosteum as a membrane separating bone and bone fluid (which may contain relatively low levels of organic phosphate) from serum. The periosteal membrane would be able to supply inorganic phosphate for hydroxyapatite formation by enzymatic hydrolysis of serum organic phosphates if and when required.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of osteoblasts and formation of mineralized bone in vitro

1982

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Summary. Periostea consisting of the osteogenic layer and the fibrous layer of the periosteum were dissected from 17-day-old embryonic chick calvariae, leaving the osteoblasts behind on bone. The dissected periostea were folded with the osteogenic cells in apposition. The explants were cultured on plasma clots for up to 6 days, during which time osteodifferentiation was observed followed by osteoid formation in between the two layers. These cultures consistently mineralized in the presence of 5 or 10 mM/3-glycerophosphate. The mineralization and osteoid formation displayed many characteristics identical with those seen in vivo. Specifically, the osteoid formed was birefringent under polarized light, the central zone of osteoid became mineralized within 24 h of formation in vitro, and a clear border between mineralized and nonmineralized osteoid suggestive of a mineralization front was present. The unmineralized osteoid at the periphery was surrounded by osteoblasts. These data suggest that physiologic mineralization of osteoid produced in vitro did occur in this system by the addition of the alkaline phosphatase substrate fl-glycerophosphate.Key words: Mineralization --Osteogenesis --Alkaline phosphatase --In vitro.To date there have been few studies showing that morphologically distinct osteoid can be formed and mineralized in vitro under cellular control and in a pattern morphologically identical to that seen in vivo. Some investigators have described culture systems in which osteoid formation occurred [1-3], whereas others have developed systems in which both osteoid formation and mineralization could be demonstrated [4][5][6][7][8][9][10][11][12]. The patterns of mineralizaSend offprint requests to H. C. Tenenbaum at the above address.of Dentistry, University of Toronto, Toronto, tion observed, however, were unpredictable and bore little similarity to the pattern of mineralization seen in vivo. Reproducible formation of mineralized bone in vitro was reported by Marvaso and Bernard [ 13], who have shown that mineralization of osteoid produced by calvarial mesenchyme occurred regularly in a pattern similar to that seen in vivo. This system, however, cannot be manipulated to obtain osteogenesis, osteodifferentiation, or mineralization at will. We describe here a system, adapted from a system first described by Nijweide and Van der Plas [2,14], that can be used to study in vitro mechanisms which promote and regulate osteodifferentiation and mineralization. Using this system, we have found that osteogenic cells from embryonic periostea differentiate in vitro into osteoblasts and subsequently produce osteoid when cultured folded with the osteogenic layers in apposition. The osteoid formed became mineralized bone when /3-glycerophosphate was added to the culture medium. The mineralized bone matrix was surrounded by a clearly distinguishable zone of nonmineralized osteoid. Materials and MethodsBriefly, periostea consisting of the osteogenic and fibrous layers (Fig. 1A) [30] were dissected from 17-day-old chick embryo...

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Section: Discussionmentioning

confidence: 99%

Differentiation of osteoblasts and formation of mineralized bone in vitro

1982

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“…In order to replicate more closely the growth and differentiation in ovo, investigators have used culture of whole embryonic bones rather than bone fragments. With such a system, the effects of numerous agents and conditions on the growth and mineralization of bone have been investigated [6]. These effects have usually been expressed quantitatively as chemical changes in the bone and/or the culture medium.…”

Section: Introductionmentioning

confidence: 99%

Growth of embryonic chick tibiae in ovo and in vitro: A scanning electron microscope study

1979

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Summary. The study describes the ultrastructure of the mineralized portion of chick tibiae from 10 days in ovo to 2 days post-hatch. At 10 days a single mineralized cylinder surrounds the diaphysis. On its outer surface columnar trabeculae join to form ridges parallel to the long axis of the bone. These ridges are covered by another cylinder and form the haversian canals. At 11 days vascular invasion of the marrow cavity occurs and resorption of the endosteal surface begins. This type of periosteal deposition and endosteal resorption is repeated during and subsequent to embryonic development.The mineralized portion of 10-day chick tibiae cultured for 2 days in modified BGJ medium was compared with 10-, 11-, and 12-day tibiae in ovo. Cultured tibiae were similar in length and calcium content to 11-day tibiae in ovo. The form of mineral deposited in ovo and in culture was the same, namely, aggregates of spherical mineral clusters. Differences in culture included the following: (a) few concentric cylinders were deposited as compared with tibiae in ovo; (b) trabeculae were not arranged in rows and ridges in culture; (c) osteocytic lacunae were restricted to bases of trabeculae rather than uniformly distributed as in ovo; and (d) the endosteal surface of tibiae in culture appeared etched.Send offprint requests to Richard M. Dillaman at the above address.

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“…For review, see Talmage20 and Ramp.2' In vitro studies have shown that the bone lining cells reduce the calcium concentration in the bone fluid bathing the mineralizing matrix by transporting calcium out of the compartment.21-24 However, the cells apparently exercise no control over the flux of phosphorus in or out of the bone fluid.21'23 The efflux of calcium reduces the Ca X Pi product in the bone fluid relative to that which exists in the general extracellular fluid, and retards mineralization in new bone. This theory is further supported by the finding that disruption of the bone membrane or metabolic inhibition of the cells of the membrane results in increased mineralization.21 '23,24 Conclusions On the basis of the observations reported here on the in vivo differences in the uptake patterns of 45Ca and 33p, and the evidence obtained in the in vitro experiments that there is a cellular limitation of flux of 45Ca, but not 33P, into newly formed enamel matrix, it seems possible that early mineralization of enamel is controlled in a manner similar to that proposed for bone. It is suggested that the relatively slow rate of mineralization in newly formed rat enamel matrix is due to a reduction in the Ca x Pi product in the fluid bathing the matrix caused, in turn, by an active movement of cal-cium away from the matrix by the secretory ameloblasts.…”

Section: Discussionmentioning

confidence: 84%

Comparison of P with Ca Distribution in Developing Rat Molar Enamel In Vivo and In Vitro

Wennberg

Bawden

1978

J Dent Res

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The distribution of 45Ca and 33P in developing rat molar enamel has been studied using freeze sectioning and autoradiographic methods. The distribution patterns of the two tracers in vivo were dissimilar. The use of in vitro methods showed that flux of 33P into enamel is not influenced by the cells of the enamel organ. It is suggested that the factors controlling movement of calcium and phosphorus into mineralizing enamel may be similar to those proposed for bone.

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Some factors affecting mineralization of bone in tissue culture

Cited by 51 publications

References 0 publications

Differentiation of osteoblasts and formation of mineralized bone in vitro

Differentiation of osteoblasts and formation of mineralized bone in vitro

Growth of embryonic chick tibiae in ovo and in vitro: A scanning electron microscope study

Comparison of P with Ca Distribution in Developing Rat Molar Enamel In Vivo and In Vitro

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