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Biomedical subjects

C G Bellows

Publications and source records attributed to C G Bellows.

At least 37 records · Page 2Linked to original sources
PubMed1990

The effects of fluoride on osteoblast progenitors in vitro.

The number of discrete, three-dimensional bone nodules formed in vitro from a class of osteoprogenitor cells present in fetal rat calvaria cell populations (RC cells) is linearly related to the number of cells plated, implying that this system functions as a colony assay for the expression of osteoprogenitor cells. To determine the effect of fluoride on the expression of these cells, primary RC cells were grown for periods of up to 21 days in alpha-MEM (minimal essential medium) containing 5-15% heat-inactivated fetal bovine serum (FBS), 50 micrograms/ml ascorbic acid, 10 mM Na beta-glycerophosphate, and NaF at concentrations from 10 microM to 5 mM. The continuous presence of NaF resulted in an increase in the number of bone nodules with maximal response occurring at 500 microM (p less than 0.001). A similar response at 500 microM NaF was observed also with regard to alkaline phosphatase activity. NaF levels up to 500 microM did not affect the growth of the mixed RC cell population, however, higher concentrations (1 mM) significantly reduced cell numbers (p less than 0.001) suggestive of cytotoxicity. Plating efficiency tests for colony formation in the presence of 0.5 to 2 mM NaF showed that the decreases in nodule formation observed at concentrations above 500 microM correlated with cytotoxicity. NaC1 at 1 mM had no effect on nodule formation, alkaline phosphatase activity, or cell growth. The results show that NaF stimulates osteoprogenitor cell number in vitro and that the maximal effect occurs at concentrations close to toxic levels.

Alkaline Phosphatase↗
PubMed1990

Determination of the capacity for proliferation and differentiation of osteoprogenitor cells in the presence and absence of dexamethasone.

Osteoprogenitor cells present in single-cell suspensions prepared from fetal rat calvaria (RC) form discrete mineralized three-dimensional bone nodules when cultured long-term in the presence of ascorbic acid and beta-glycerophosphate. These cells (CFU-O) constitute less than 1% of the total cell population under standard culture conditions and their number is increased in the presence of dexamethasone. Using the formation of the bone nodule as a marker for CFU-O, we have now analyzed the proliferation and differentiation capacity of these CFU-O by redistribution and continuous subculture experiments in the presence and absence of dexamethasone. Cell redistribution experiments showed no increase in nodule number after one population doubling with either treatment. After 5.4 population doublings of the entire RC population, nodule number increased up to 2.0-fold in control cultures and 4.5-fold in cultures containing 10 nM dexamethasone. Continuous subculture experiments in which cultures were split 1:3 every 3 day for up to seven subcultures showed that nodule number decreased in parallel with the split ratio in the absence of dexamethasone, while with dexamethasone nodule number was elevated above the number present in primary cultures for 1 or 2 subcultures after which nodule number decreased with the split ratio. Bone nodules were present for up to 18 population doublings. Measurements of nodule area by automated image analysis showed that dexamethasone increased nodule size and that nodule size decreased from primary to 1st to 2nd subculture with or without dexamethasone. The data suggest that dexamethasone selectively stimulates the proliferation of osteoprogenitor cells and that these progenitor cells have a limited capacity for generating daughter cells capable of expressing the bone phenotype.

Animals↗
PubMed1990

Parathyroid hormone reversibly suppresses the differentiation of osteoprogenitor cells into functional osteoblasts.

The effects of PTH on osteoprogenitor cell differentiation have been analyzed by quantifying its effects on bone nodule formation in an in vitro assay. Fetal rat calvaria cells were plated at 3 x 10(4) cells/35-mm dish, and cultures were maintained for 17-23 days in alpha-Minimal Essential Medium containing ascorbic acid, Na beta-glycerophosphate, and 10% fetal bovine serum. Continuous exposure to PTH at concentrations from 1 pM to 1 nM (2 x 10(-5) to 2 x 10(-2) IU/ml) caused a dose-dependent inhibition of bone nodule formation. Half-maximal inhibition occurred at 0.05 nM, and total inhibition at 1 nM, concentrations much lower than those required to elicit a significant cAMP response in rat calvaria cells. PTH at the concentrations used did not affect cell growth or saturation density. While continuous exposure to 1 nM PTH eliminated bone nodule formation, a single 48-h pulse administered at any time during the 17-day culture period had no effect. When 1 nM PTH was added on day 1 and removed at different times during the culture period, a time-related release from inhibition was observed. Cultures exposed to 1 nM PTH until nodules had developed in the corresponding control cultures and then switched to medium without added PTH rapidly formed clusters of differentiated osteoblasts and nodules within 3 days. PTH added at different times during the culture period and present continuously there-after suppressed formation of new nodules, the magnitude of the effect being a function of the duration of exposure. The results show that PTH at physiological concentrations is a potent suppressor of osteoblast differentiation and that its effect occurs at a late stage in the differentiation of osteoprogenitor cells, probably preventing differentiation of preosteoblasts into osteoblasts.

Animals↗
PubMed1989

Effects of transforming growth factor beta and epidermal growth factor on cell proliferation and the formation of bone nodules in isolated fetal rat calvaria cells.

When cells enzymatically isolated from fetal rat calvaria (RC cells) are cultured in vitro in the presence of ascorbic acid and Na beta-glycerophosphate, discrete three-dimensional nodules form with the histologic, immunohistochemical, and ultrastructural characteristics of bone (Bellows et al; Calcified Tissue International 38:143-154, 1986; Bhargava et al., Bone, 9:155-163, 1988). Quantitation of the number of bone nodules that forms provides a colony assay for osteoprogenitor cells present in the RC population (Bellows and Aubin, Develop. Biol., 133:8-13, 1989). Continuous culture with either epidermal growth factor (EGF) or transforming growth factor beta (TGF-beta) results in dose-dependent inhibition of bone nodule formation; however, the former causes increased proliferation and saturation density, while the latter reduces both parameters. Addition of EGF (48 h pulse, 2-200 ng/ml) to RC cells at day 1 after plating results in increased proliferation and population saturation density and an increased number of bone nodules formed. Similar pulses at confluence and in postconfluent multilayered cultures when nodules first begin forming (approx. day 11) inhibited bone nodule formation and resulted in a smaller stimulation of cell proliferation. Forty-eight hour pulses of TGF-beta (0.01-1 ng/ml) reduced bone nodule formation and proliferation at all times examined, with pulses on day 1 causing maximum inhibition. The effects of pulses with TGF-beta and EGF on inhibition of nodule formation are independent of the presence of serum in the culture medium during the pulse. The data suggest that whereas EGF can either stimulate or inhibit the formation of bone nodules depending upon the time and duration of exposure, TGF-B inhibits bone nodule formation under all conditions tested. Moreover, these effects on osteoprogenitor cell differentiation do not always correlate with the effects of the growth factors on RC cell proliferation.

Animals↗
PubMed1989

Effects of dexamethasone on expression and maintenance of cartilage in serum-containing cultures of calvaria cells.

The effects of dexamethasone on the ability of cells enzymatically isolated from 21-day fetal rat calvaria to produce cartilage in vitro has been investigated. Primary cultures of single-cell suspensions of rat calvaria were grown for up to 28 days in vitro in alpha-minimal essential medium containing 15% fetal bovine serum, 50 micrograms/ml ascorbic acid, 10 mM Na beta-glycerophosphate and dexamethasone at concentrations of 1 microM to 1 nM. Two types of nodules were present in dexamethasone-containing cultures. One has been characterized previously as bone (Bellows et al. 1986). The second morphologically resembled hyaline cartilage, possessed a strong Alcian blue-positive matrix and contained type-II, but not type-I, collagen. Both bone and cartilaginous nodules were spatially distinct and developed in isolation from each other. Cartilaginous nodules were found in the highest number at a dexamethasone concentration of 100 nM. Time-course experiments revealed that while the number of bone nodules increased continuously at least to day 28, the number of cartilaginous nodules remained constant after cultures had reached confluency. When cells were isolated separately from frontal and parietal bones and sutural regions, the greatest number of cartilaginous nodules developed from parietal bones. Since 21-day fetal rat calvaria contains 2 distinct patches of cartilage at the periphery of the parietal bones, it seems likely that this cartilaginous tissue is the origin of the cartilage cells. The results demonstrate that cultures of rat calvaria cells contain chondrocytes and possibly chondroprogenitor cells that are distinct from osteoprogenitors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Determination of numbers of osteoprogenitors present in isolated fetal rat calvaria cells in vitro.

When maintained in long-term cell culture in the presence of ascorbic acid and organic phosphate, single cell suspensions isolated from fetal rat calvaria form discrete, three-dimensional bone nodules. We have used limiting dilution analysis in microtiter wells to determine the number of osteoprogenitor cells expressing the capacity to form bone in the isolated mixed population, to examine the possibility of cooperativity among cell types in bone nodule formation, and to determine the effects of dexamethasone on osteoprogenitor cells. Cells plated at very low densities and screened for the presence or absence of bone nodules revealed a linear relationship (r = -00.997) between the number of cells plated and the number of bone nodules formed. The complete limiting dilution analyses showed that 1 of every 335 plated cells (0.30% of the cell population) has the capacity to form a bone nodule under standard culture conditions and when the actual numbers of nodules were quantitated from the same plated cell populations the ratio of nodules formed to plated cells was similar. Comparison of data from 13 different isolates of cells in which cells were plated into 35-mm dishes and number of nodules were determined indicated a mean +/- 95% confidence interval of one nodule for every 301 +/- 61 plated cells, consistent with the data obtained from the limiting dilution experiments. Dexamethasone increased the number of bone-forming cells to 1 in 225 cells, in contrast to 1 in 340 cells in the same population grown without added dexamethasone. The results suggest that approximately 0.30% of the cells in isolated rat calvaria populations are osteoprogenitor cells, that one osteoprogenitor cell gives rise to one bone nodule, that cooperativity between different cells in vitro is not necessary for bone formation, and that dexamethasone stimulates the expression of osteoprogenitor cells.

Animals↗
PubMed1988

Ultrastructural analysis of bone nodules formed in vitro by isolated fetal rat calvaria cells.

When cells enzymatically digested from 21 d fetal rat calvaria are grown in ascorbic acid and Na beta-glycerophosphate, they form discrete three-dimensional nodular structures with the histological and immunohistochemical appearance of woven bone. The present investigation was undertaken to verify that bone-like features were identifiable at the ultrastructural level. The nodules formed on top of a fibroblast-like multilayer of cells. The upper surface of the nodules was lined by a continuous layer of cuboidal osteoblastic cells often seen to be joined by adherens junctions. Numerous microvilli, membrane protrusions, and coated pits could be seen on the upper surface of these cells, their cytoplasm contained prominent RER and Golgi membranes, and processes extended from their lower surfaces into a dense, highly organized collagenous matrix. Some osteocyte-like cells were completely embedded within this matrix; they also displayed RER and prominent processes which extended through the matrix and often made both adherens and gap junctional contacts with the processes of other cells. The fibroblastic cells not participating in nodule formation were surrounded by a less dense collagenous matrix and, in contrast to the matrix of the nodules, it did not mineralize. An unmineralized osteoid-like layer was seen directly below the cuboidal top layer of cells. A mineralization front was detectable below this in which small, discrete structures resembling matrix vesicles and feathery mineral crystals were evident and frequently associated with the collagen fibrils. More heavily mineralized areas were seen further into the nodule. Electron microprobe and electron and X-ray diffraction analysis confirmed the mineral to be hydroxyapatite.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Biphasic effects of epidermal growth factor on bone nodule formation by isolated rat calvaria cells in vitro.

Osteoblast-like cells isolated from fetal rat calvaria (RC) form three-dimensional nodules in vitro having the morphological and immunohistochemical characteristics of bone. When administered continually over 21 days of culture, EGF caused a dose-related inhibition of nodule formation, as well as a reduction in the proportion of mineralized nodules, at concentrations between 10(-12) to 10(-7) M. The same conditions caused an increase in both cell proliferation and saturation density in the cultures, suggesting that decreased nodule formation was not the result of general cell toxicity. Inhibitory concentrations of EGF caused a reversible alteration in cell shape, although changes in protein synthesis were not detectable. Indomethacin (5 X 10(-7) M) did not affect either nodule formation or its inhibition by EGF. When cultures were pretreated for various time periods with 10(-8) M EGF followed by its removal and continued culture in supplemented medium for 21 days, the number of bone nodules formed with brief exposures to EGF (4 hr up to 48 hr) was increased compared to the numbers formed in supplemented medium alone, whereas exposures to EGF of 4 days or longer decreased the number of nodules formed. These data indicate that EGF can either increase or decrease the osteogenic potential of RC cells in vitro depending on the duration of exposure to the factor.

Animals↗
PubMed1987

Physiological concentrations of glucocorticoids stimulate formation of bone nodules from isolated rat calvaria cells in vitro.

Isolated rat calvaria cells plated at low density in medium supplemented with ascorbic acid and organic phosphate form discrete three-dimensional mineralized nodules having the characteristics of bone. We have studied the effects of glucocorticoids on the formation of bone nodules by these cell populations. Cells isolated from 21-day-old fetal rat calvaria were maintained in vitro for up to 27 days. Dexamethasone (Dex) induced a dose-related increase in the number of nodules formed, with a peak at 10 nM and a half-maximal response at about 1 nM. Dex (10 nM) also significantly increased the size of bone nodules formed (P less than 0.002). High concentrations of Dex (1 microM) did not increase nodule number. In cells in primary culture maintained in medium containing 10 nM Dex, the increase in nodule number was 50-100% over the control value. The effect of Dex was much greater in first subculture cells, where the number of nodules was 600-800% higher than the control value. Dishes collected and quantitated from 12-27 days showed that nodule formation ceased between 15 and 18 days in cultures without Dex, whereas in the presence of Dex the number of nodules increased up to 27 days. Addition of 10 nM Dex only during specific periods resulted in significantly more nodules than in control cultures, but significantly fewer nodules than in cultures constantly exposed to Dex. Cell population doubling times during log phase growth were unaltered, but a significant increase in saturation density (P less than 0.001) was observed with 10 nM Dex. Hydrocortisone also caused an increase in the number of nodules formed, with a maximal effect of 50 nM and a half-maximal response at 8 nM. The results indicate that physiological levels of glucocorticoids stimulate bone nodule formation in long term cell culture by increasing the number of cells forming bone nodules and that maximization of the stimulatory effect of glucocorticoids on bone formation may require constant exposure to low levels of the hormone.

Animals↗
PubMed1986

The effects of platelet-derived transforming growth factor beta on normal human diploid gingival fibroblasts.

Studies of the effects of transforming growth factor (TGF) beta on normal human diploid gingival fibroblasts (HGF) have been carried out to determine possible physiological effects of this growth factor. Responses distinctly different from those characterized using established cell lines were observed. Whether alone, or in combination with EGF (2.5 ng/ml), human platelet-derived TGF-beta (0.1 ng/ml or 1.0 ng/ml) did not induce anchorage-independent growth of HGFs in soft agar assays. However, TGF-beta with EGF acted synergistically in promoting a 1.8-fold increase in anchorage-dependent proliferation of quiescent HGFs. At the same concentrations TGF-beta alone stimulated the incorporation of [35S]methionine into both cellular (cell-layer) and matrix (medium) proteins by as much as 3-fold and 1.7-fold respectively. Densitometric analysis of fluorographs of radiolabeled media proteins separated by SDS-PAGE revealed that the TGF-beta-stimulated protein synthesis was selective. However, synthesis of collagen, the major protein synthesized and secreted by HGFs, was stimulated by TGF-beta to the same extent as the average secreted protein. Protein synthesis and cell proliferation were significantly greater in subconfluent cells compared to confluent and multilayered cells. These effects are likely to reflect physiological activity of platelet-derived TGF-beta which may act to promote the wound healing response.

Amino Acids↗
PubMed1986

Phenotypic differences in subclones and long-term cultures of clonally derived rat bone cell lines.

Previous studies with clonally derived populations of cells have shown that cells released from embryonic rat calvaria by enzymatic digestion are heterogeneous with respect to their hormone responsiveness, morphology, and production of matrix components [Aubin JE et al; J. Cell Biol 92:452, 1982]. Several of these clonal populations have been used to study the effects of long-term culture and inter- and intraclonal cell heterogeneity. During continuous subculture, marked changes in collagen synthesis were observed in two clonal populations. Both of these clones were originally responsive to parathyroid hormone (PTH) and synthesized primarily type I collagen with small amounts of type III and V collagens, although one clone (RCJ 3.2) had a fibroblastic morphology whereas the second clone (RCB 2.2) displayed a more polygonal shape. Following routine subculture over 3 yr, clone RCB 2.2 was found to synthesize exclusively alpha 1(I)-trimer and not other interstitial collagens. When the same cells were maintained at confluence for 1-2 wk, however, they also synthesized type III collagen. Whereas RCJ 3.2 did not show such dramatic changes in collagen synthesis after long-term subculture, two subclones derived from RCJ 3.2 were found to synthesize almost exclusively either type III collagen (RCJ 3.2.4.1) or type V collagen (RCJ 3.2.4.4). Immunocytochemical staining indicated that both subpopulations also produced type IV collagen, laminin, and basement membrane proteoglycan, proteins that are typically synthesized by epithelial cells. The differences in collagen expression by the various clonal cell populations were accompanied by qualitative and quantitative differences in other secreted proteins and differences in cell morphology. The results demonstrate both the inter- and intraclonal heterogeneity of connective tissue cells and their diverse potentiality with respect to extracellular matrix synthesis.

Animals↗
PubMed1986

Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations.

Single-cell suspensions obtained from sequential enzymatic digestions of fetal rat calvaria were grown in long-term culture in the presence of ascorbic acid, Na beta-glycerophosphate, and dexamethasone to determine the capacity of these populations to form mineralized bone. In cultures of osteoblastlike cells grown in the presence of ascorbic acid and beta-glycerophosphate or ascorbic acid alone, three-dimensional nodules (approximately 75 micron thick) covered by polygonal cells resembling osteoblasts could be detected 3 days after confluency. The nodules became macroscopic (up to 3 mm in diameter) after a further 3-4 days. Only in the presence of organic phosphate did they mineralize. Nodules did not develop without ascorbic acid in the medium. Dexamethasone caused a significant increase in the number of nodules. Histologically, nodules resembled woven bone and the cells covering the nodules stained strongly for alkaline phosphatase. Immunolabeling with specific antibodies demonstrated intense staining for type I collagen that was mineral-associated, a weaker staining for type III collagen and osteonectin, and undetectable staining for type II collagen. Nodules did not develop from population I and the number of nodules formed by populations II-V bore a linear relationship to the number of cells plated (r = .99). The results indicate that enzymatically released calvaria cells can form mineralized bone nodules in vitro in the presence of ascorbic acid and organic phosphate.

Animals↗
PubMed1983

An in-vitro model for tooth eruption utilizing periodontal ligament fibroblasts and collagen lattices.

An in-vitro model that simulates tooth eruption was devised to determine the possible role of periodontal ligament fibroblasts (PLF) in providing a major force for eruption. The model consisted of agarose-coated wells in multiwell tissue-culture plates to which wire mesh was attached at the rim and a slice of demineralized root was attached to the floor. When the wells were filled with a collagen lattice containing PLF, the lattice contracted and elevated the root slice. The root slice in models that were unattached at the rim was not elevated. Cultures without cells did not contract and root slices were not elevated. Elevation was inhibited by cytochalasin D and Colcemid. Histologically, cultures that had elevated showed orientated cells and collagen fibres. The results suggest that PLF may be capable of generating forces that are sufficient to produce tooth eruption.

Animals↗
PubMed1982

Association between tension and orientation of periodontal ligament fibroblasts and exogenous collagen fibres in collagen gels in vitro.

The relationship between the development of tension in sheets of fibroblasts and the orientation of these cells and collagen fibres in collagen gels was examined. Cell-containing, three-dimensional collagen gels were established in agarose-coated Epon dies measuring 10 mm X 4 mm X 4 mm, to which pieces of demineralized tooth and bone had been attached at opposite ends. Contraction of the gel into an opaque structure suspended between the two particles occurred over 24 h and resulted in concave upper and lateral surfaces and a flat to slightly concave lower surface. Initial orientation of the fibres along the tooth-bone axis was followed by similar orientation of the cells. Gels cast without cells exhibited no change in dimensions. Release of the tooth particle after 12 or 24 h of incubation led to shortening of the contracted gels 5 min following release. This shortening was significantly greater (P less than 0.001) than that of uncontracted or slightly contracted gels (1 h and 3 h incubation). Gels attached at one end only compacted around the site of attachment but did not show orientation of cells or fibres. Gels containing colcemid or cytochalasin D were only slightly compacted and did not develop tension. Collagen fibres, but not cell in colcemid-containing gels, showed some alignment; neither were aligned in the presence of cytochalasin D. These data suggest that both microtubules and microfilaments are necessary for alignment of cells and the establishment of tension between two points of attachment in collagen gels. Furthermore, they lend support to our previously advanced hypothesis that the development of tension between two points can result in the orientation of the cells along an axis connecting the points of attachment. This could provide a mechanism for the development of oriented fibre systems in vivo.

Animals↗
PubMed1981

Contraction and organization of collagen gels by cells cultured from periodontal ligament, gingiva and bone suggest functional differences between cell types.

Monkey periodontal ligament fibroblasts (MPLF cells), human gingival fibroblasts (HGF cells), rat embryonic calvaria cells (REC cells), porcine periodontal ligament epithelial cells (PPLE cells) and rat osteosarcoma 17/2 cells (ROS cells) were incorporated into 3-dimensional collagen gels plated in 60 mm Petri dishes in order: first, to measure the capacity of these cell types to contract; second, to investigate cell-collagen and intercellular relationships during contraction; and third, to define the cellular contribution to tissue contraction in an in vitro system. Measurements at times up to 72 h on 3 ml gels containing 5 x 10(5) cells and with a collagen concentration of 1.20 mg/ml showed that MPLF cells contracted the gels at a significantly greater rate (P less than 0.001) than did the other cell types. In addition, contraction started sooner and was of greater extent than with the other cells. HGF cells contracted the gels more rapidly than REC and PPLE cells, while ROS cells caused no contraction. Several stages of gel compaction could be defined: (1) the attachment of cells to collagen; (2) cellular spreading within the collagen fibre matrix; (3) organization and alignment of collagen fibres by cell processes; (4) cell migration; (5) establishment of intercellular contacts; and (6) the development of a cellular reticular arrangement within the gel and the extension of this arrangement into a 3-dimensional, tissue-like, honeycomb network. Electron microscopic observations on 0.1 ml gels containing MPLF cells showed that, in the early contractile phase, numerous cell processes attached to or enclosed collagen fibrils. These processes contained microfilamentous material and few organelles. In compacted gels, the cells contained an increased amount of distended rough endoplasmic reticulum and Golgi membranes. Since MPLF cells have the capacity for vigorous contraction of the collagen gels and since they develop a reticular, 3-dimensional structure in compacted gels that is reminiscent of the relationship of periodontal ligament fibroblasts to collagen fibres in vivo, it is suggested that they could provide the major force necessary for tooth eruption in vivo. This system also provides a well-defined in vitro model to study the sequential stages that occur during contraction processes.

Animals↗
PubMed1980

Orientation of calvaria and periodontal ligament cells in vitro by pairs of demineralized dentine particles.

Rat calvaria cells (RC cells) and monkey periodontal ligament fibroblasts (MPL cells) were cultured for 4-6 days with demineralized dentine, acid-washed glass particles or untreated glass particles to determine whether the presence of these particles can affect the orientation of the cells. Encapsulation of dentine particles began as early as the first day in culture and occasional cells were attached to, and oriented between adjacent particles. Multilayers of cells formed along the periphery of the dentine particles and the more superficial of these cells projected outwards in a radial fashion. Oriented cell sheets were evident between dentine particles after 3 days in culture. Finally, the cell sheets tore away from the surface of the culture fish to give rise to thick multilayered cellular bridges between dentine particles. The nuclear orientation index (ratio of L/W) for each RC cell lying between 15 randomly selected pairs of each particle type in 4-day cultures was measured along 2 axes; the length represented by the shortest distance between the 2 particles and the width perpendicular to this axis. The nuclear orientation index was 1.52 for cells between dentine, 1.13 between acid-washed glass and 1.06 between untreated glass. Control measurements of cells associated with single particles were 1.06, 1.09 and 1.06 respectively. The frequency of cellular orientation occurring between dentine particles was significantly greater than that occurring between acid-washed glass particles (chi 2 > 0.001) which in turn was significantly greater than that for untreated glass particles (chi 2 > 0.001). The observations and results suggest that the development and maintenance of the observed cell orientation depends upon development and maintenance in the cell sheet of tensional forces oriented parallel to the axis between the dentine particles, and that this is related directly to the capacity of the cells to attach to the particles.

Animals↗