Net proton influx into bone during metabolic, but not respiratory, acidosis

1988 ◽  
Vol 254 (3) ◽  
pp. F306-F310 ◽  
Author(s):  
D. A. Bushinsky
Keyword(s):  
Metabolic Acidosis ◽  
Bone Mineral ◽  
Respiratory Acidosis ◽  
Mineral Dissolution ◽  
Calcium Efflux ◽  
Bicarbonate Concentration ◽  
Medium Ph ◽  
Acute Metabolic Acidosis ◽  
Initial Medium

During acute metabolic acidosis there is a net influx of protons into bone, decreasing the elevated proton concentration. Whether there is an influx of protons into bone during acute respiratory acidosis is not known. To determine the effect of respiratory acidosis on net proton flux (JH) relative to bone, we compared JH from neonatal mouse calvariae incubated for 3 h in medium acidified by an increase in PCO2 (respiratory acidosis) with that from calvariae incubated in medium acidified to the same extent by a decrease in bicarbonate concentration (metabolic acidosis). The initial medium pH with respiratory acidosis was not different from that with metabolic acidosis (7.108 +/- 0.005 vs. 7.091 +/- 0.007, respectively, P = NS). During respiratory acidosis there was no JH from bone relative to the medium (JH = 236 +/- 93 neq.bone-1.3h-1, P = NS vs. 0); however, during metabolic acidosis there was net proton influx from the medium into bone (JH = -703 +/- 108, P less than 0.05 vs. 0, P less than 0.001 vs. respiratory acidosis). There was less calcium efflux from bone during respiratory than during metabolic acidosis (JCa = 68 +/- 6 nmol.bone-1.3 h-1 vs. 100 +/- 9, respectively, P less than 0.001). There is a net influx of protons into bone in vitro during acute metabolic, but not during acute respiratory, acidosis. The smaller calcium efflux during respiratory acidosis may indicate less net bone mineral dissolution and thus less buffer release into the medium.

Net calcium efflux from live bone during chronic metabolic, but not respiratory, acidosis

1989 ◽  
Vol 256 (5) ◽  
pp. F836-F842 ◽  
Author(s):  
D. A. Bushinsky
Keyword(s):  
Metabolic Acidosis ◽  
Bone Cells ◽  
Respiratory Acidosis ◽  
Neonatal Mouse ◽  
Mineral Dissolution ◽  
Calcium Flux ◽  
Calcium Efflux ◽  
Physicochemical Factors ◽  
Acute Metabolic Acidosis

In vivo chronic metabolic acidosis induces bone mineral dissolution. Whether the dissolution is due to alterations in physicochemical factors alone, as in acute metabolic acidosis, or requires participation of bone cells is not clear. The effect of chronic respiratory acidosis on bone has also not been established. To determine the effects of chronic metabolic and respiratory acidosis on net calcium flux from bone, we cultured live and dead neonatal mouse calvariae for 99 h in control medium or in medium acidified (pH approximately equal to 7.1) either by lowering the bicarbonate concentration (Met) or by increasing the PCO2 (Resp). We measured net calcium flux (JCa) over 0-48, 48-96, and 96-99 h. Over the first 48 h, there was greater net calcium efflux from live and dead Met than from both Resp groups. All four acidic groups had greater net calcium efflux than controls. Over the last 51 h of the chronic 99 h culture, there was net calcium efflux only from live Met (JCa = 285 +/- 129 nmol.bone-1.3 h-1) and not from any of the other groups (live control, JCa = -183 +/- 24; live Resp, JCa = -110 +/- 22; dead control, JCa = -256 +/- 12; dead Met, JCa = 11 +/- 78; dead Resp, JCa = -27 +/- 47; each P less than 0.02 vs. live Met). There is net calcium efflux from live cultured neonatal mouse calvariae during chronic metabolic, but not respiratory, acidosis. During chronic acidosis, decreased medium bicarbonate, and not just a fall in pH, is necessary to enhance net calcium efflux from live bone.

Decreased bone carbonate content in response to metabolic, but not respiratory, acidosis

1993 ◽  
Vol 265 (4) ◽  
pp. F530-F536 ◽  
Author(s):  
D. A. Bushinsky ◽  
B. C. Lam ◽  
R. Nespeca ◽  
N. E. Sessler ◽  
M. D. Grynpas
Keyword(s):  
Metabolic Acidosis ◽  
Respiratory Acidosis ◽  
Calcium Flux ◽  
Carbonate Content ◽  
Carbonate Dissolution ◽  
Bicarbonate Concentration ◽  
Medium Ph ◽  
Carbonated Apatite ◽  
Time Periods

In vitro cultured neonatal mouse calvariae release calcium and buffer the medium proton concentration in response to a decrease in the medium pH caused by a reduction in bicarbonate concentration ([HCO3-]), a model of metabolic acidosis, but not to an equivalent decrease in pH caused by an increase in the partial pressure of carbon dioxide (PCO2), a model of respiratory acidosis. We have postulated that the medium is in equilibrium with the carbonated apatite in bone. To determine whether bone carbonate is depleted during models of acidosis, we cultured calvariae in control medium (pH approximately 7.4, PCO2 approximately 43, [HCO3-] approximately 26) or in medium in which the pH was equivalently reduced by either a decrease in [HCO3-] (metabolic acidosis, pH approximately 7.1, [HCO3-] approximately 13) or an increase in PCO2 (respiratory acidosis, pH approximately 7.1, PCO2 approximately 86) and determined net calcium flux (JCa) and bone carbonate content. We found that compared with control, after 3, 24, and 48 h there was a decrease in bone carbonate content during metabolic but not during respiratory acidosis. Compared with control, at 3 h JCa increased with both respiratory and metabolic acidosis; however, at 24 and 48 h JCa increased only with metabolic acidosis. JCa was correlated inversely with percent bone carbonate content in control and metabolic acidosis at all time periods studied (r = -0.809, n = 23, P < 0.001). Thus a model of metabolic acidosis appears to increase JCa from bone, perhaps due to the low [HCO3-] inducing bone carbonate dissolution.(ABSTRACT TRUNCATED AT 250 WORDS)

Metabolic alkalosis decreases bone calcium efflux by suppressing osteoclasts and stimulating osteoblasts

1996 ◽  
Vol 271 (1) ◽  
pp. F216-F222 ◽  
Author(s):  
D. A. Bushinsky
Keyword(s):  
Bone Mineral ◽  
Metabolic Alkalosis ◽  
Collagen Synthesis ◽  
Cell Function ◽  
Urinary Calcium ◽  
Calcium Excretion ◽  
Calcium Efflux ◽  
Medium Ph ◽  
Bone Calcium

In vivo and in vitro evidence indicates that metabolic acidosis, which may occur prior to complete excretion of end products of metabolism, increases urinary calcium excretion. The additional urinary calcium is almost certainly derived from bone mineral. Neutralization of this daily acid load, through the provision of base, decreases calcium excretion, suggesting that alkali may influence bone calcium accretion. To determine whether metabolic alkalosis alters net calcium efflux (JCa+) from bone and bone cell function, we cultured neonatal mouse calvariae for 48 h in either control medium (pH approximately equal to 7.4, [HCO3-] approximately equal to 24), medium simulating mild alkalosis (pH approximately equal to 7.5, [HCO3-] approximately equal to 31), or severe alkalosis (pH approximately equal to 7.6, [HCO3-] approximately equal to 39) and measured JCa+ and the release of osteoclastic beta-glucuronidase and osteoblastic collagen synthesis. Compared with control, metabolic alkalosis caused a progressive decrease in JCa+, which was correlated inversely with initial medium pH (pHi). Alkalosis caused a decrease in osteoclastic beta-glucuronidase release, which was correlated inversely with pHi and directly with JCa+. Alkalosis also caused an increase in osteoblastic collagen synthesis, which was correlated directly with pHi and inversely with JCa+. There was a strong inverse correlation between the effects alkalosis on osteoclastic beta-glucuronidase release and osteoblastic collagen synthesis. Thus metabolic alkalosis decreases JCa+ from bone, at least in part, by decreasing osteoclastic resorption and increasing osteoblastic formation. These results suggest that the provision of base to neutralize endogenous acid production may improve bone mineral accretion.

Mechanism of proton-induced bone calcium release: calcium carbonate-dissolution

1987 ◽  
Vol 253 (5) ◽  
pp. F998-F1005 ◽  
Author(s):  
D. A. Bushinsky ◽  
R. J. Lechleider
Keyword(s):  
Metabolic Acidosis ◽  
Calcium Carbonate ◽  
Bone Mineral ◽  
Calcium Release ◽  
Calcium Efflux ◽  
Carbonate Phase ◽  
Bone Calcium ◽  
Calcium Carbonate Dissolution

Protons are buffered and calcium is released by bone during metabolic acidosis. Incubation of neonatal mouse calvariae in acid medium causes net calcium efflux from bone and net proton influx into bone, just as metabolic acidosis does in vivo. To determine whether the calcium carbonate phase of bone mineral is solubilized with increasing proton concentrations, we cultured calvariae for 3 h in medium in which the saturation was varied by changing pH or calcium and phosphate concentrations. We determined the driving force for crystallization by calculating the Gibbs free energy of formation (DG). With alteration of the medium pH, calcium carbonate entry or loss from bone varied linearly with the initial DG for medium calcium carbonate (r = -0.745, n = 41, P less than 0.001) as it did with alteration of the medium calcium and phosphate (r = -0.665, n = 118, P less than 0.001). There was dissolution of calcium carbonate into medium that was unsaturated with respect to calcium carbonate, net flux ceased at saturation, and calcium carbonate entered bone from supersaturated medium, indicating that the medium is in equilibrium with the calcium carbonate phase of bone mineral. Neither the mineral phase brushite nor apatite was in equilibrium with the medium. These observations indicate that in vitro, acute proton-induced calcium efflux is due to dissolution of bone calcium carbonate.

Ammoniagenesis by the Isolated Perfused Rat Kidney: The Critical Role of Urinary Acidification

1979 ◽  
Vol 56 (4) ◽  
pp. 353-364 ◽  
Author(s):  
R. L. Tannen ◽  
B. D. Ross
Keyword(s):  
Metabolic Acidosis ◽  
Rat Kidney ◽  
Critical Role ◽  
Sodium Reabsorption ◽  
Ammonia Production ◽  
Bicarbonate Concentration ◽  
Urine Ph ◽  
Average Value ◽  
Acute Metabolic Acidosis

1. The effect of metabolic acidosis simulated in vitro on ammoniagenesis was investigated by using the isolated kidney of the rat perfused with an albumin Krebs—Henseleit medium containing glutamine and glucose. 2. Addition of HCl to a perfusate of normal bicarbonate concentration resulted in a prompt increase in urine flow rate, decrease in fractional sodium reabsorption and decrease in urine pH. 3. A minimum urine pH as low as 5·15 was achieved, with an average value of 5·92, indicating that this preparation has the capacity to acidify normally. 4. In contrast with studies in vitro with other preparations, with the functional perfused kidney a diminution in perfusate bicarbonate concentration resulted in a prompt increase in ammonia production, which was strikingly correlated with the decrease in urine pH. 5. The increase in ammonia production was diminished in studies carried out with a non-urinating kidney, in comparison with those that exhibited significant urine acidification. 6. These data suggest that a decrease in urine pH with trapping of ammonia in the urine may be a critical stimulus for increased ammonia production in acute metabolic acidosis.

Effects of medium pH on duodenal and ileal calcium active transport in the rat

1986 ◽  
Vol 251 (5) ◽  
pp. G695-G700 ◽  
Author(s):  
M. J. Favus ◽  
D. A. Bushinsky ◽  
F. L. Coe
Keyword(s):  
Metabolic Acidosis ◽  
Active Transport ◽  
Direct Effect ◽  
Respiratory Acidosis ◽  
Medium Ph ◽  
Effect Of Ph ◽  
Ca Transport ◽  
Normal Chow

To study the effect of pH on Ca active transport in vitro pH was varied from 7.1 to 7.7, and bidirectional transmural Ca fluxes were measured under short-circuited conditions across duodenum or ileum from rats fed either normal chow (NCD, 1.2% Ca) or low Ca (LCD, 0.002% Ca). Duodenum and ileum from rats fed LCD actively absorbed calcium at medium pH 7.4. Reduction in mucosal and serosal medium bicarbonate from 25 to 10 mM (pH 7.4 to 7.1) decreased duodenal net Ca absorption (Jnet) from 121 +/- 32 to 39 +/- 9 nmol X cm-2 X h-1 (P less than 0.02) and ileal Jnet from 74 +/- 13 to 22 +/- 6 (P less than 0.01). The decline in duodenal and ileal Ca Jnet was due to a decrease in the mucosal-to-serosal flux (Jm----s). Raising medium pH from 7.4 to 7.7 by increasing bicarbonate from 25 to 50 mM did not alter Ca Jm----s, serosal-to-mucosal flux (Js----m), or Jnet X Ca fluxes across ileum from rats fed NCD demonstrated net secretion. LCD ileal Jm----s was unaltered when pH was reduced from 7.4 to 7.1 by increasing the PCO2 content of the buffer from 30 to 81 mmHg while maintaining bicarbonate at 24 mM. The results indicate that in vitro conditions that simulate metabolic acidosis (low bicarbonate and pH, normal PCO2) inhibit 1,25D-mediate calcium Jm----s, whereas conditions that simulate respiratory acidosis have no effect on Ca fluxes. The present studies suggest that decreases in calcium Jm----s is by a primary alteration in transport of other ions rather than direct effect on Ca transport.

Greater unidirectional calcium efflux from bone during metabolic, compared with respiratory, acidosis

1992 ◽  
Vol 262 (3) ◽  
pp. F425-F431 ◽  
Author(s):  
D. A. Bushinsky ◽  
N. E. Sessler ◽  
N. S. Krieger
Keyword(s):  
Driving Force ◽  
Respiratory Acidosis ◽  
Mineral Dissolution ◽  
Calcium Flux ◽  
Calcium Efflux ◽  
Carbonated Apatite ◽  
Acid Base Status ◽  
45Ca Efflux ◽  
45Ca Release

There is a smaller net calcium efflux from bone in vitro during respiratory (increased PCO2) than metabolic (decreased [HCO3-] acidosis. This could be due to the elevated PCO2, which would lessen the driving force for mineral dissolution and increase the driving force for mineralization with respect to carbonated apatite in the bone mineral. To test this hypothesis, we injected neonatal mice with 45Ca and dissected the radiolabeled calvariae 24 h later. The live calvariae were then cultured for 24 h under conditions simulating respiratory acidosis (Resp, pH = 7.225 +/- 0.003, PCO2 = 87.5 +/- 0.1 mmHg), severe respiratory acidosis (SResp, pH = 7.072 +/- 0.004, PCO2 = 103.0 +/- 0.5 mmHg), metabolic acidosis (Met, pH = 7.212 +/- 0.003, HCO3- = 15.5 +/- 0.1 meq/l), or normal acid-base status (Ctl, pH = 7.452 +/- 0.003, PCO2 = 40.0 +/- 0.2 mmHg, HCO3- = 27.8 +/- 0.2 meq/l) and bidirectional net calcium flux (JCa) and unidirectional 45Ca release were determined. There was greater JCa from bone during Met than Resp, and JCa was not different from Met during SResp despite the latter having a significantly lower pH. There was greater unidirectional 45Ca release from bone during Met than Resp, SResp, or Ctl. There was a similar direct correlation between JCa and 45Ca efflux in the respiratory and metabolic groups. However, when calvarial osteoclast activity was inhibited with calcitonin,although there was again greater JCa and 45Ca release with a metabolic compared with respiratory acidosis, there was a greater proportion of 45Ca release than JCa from bone.(ABSTRACT TRUNCATED AT 250 WORDS)

Metabolic, but not respiratory, acidosis increases bone PGE2 levels and calcium release

2001 ◽  
Vol 281 (6) ◽  
pp. F1058-F1066 ◽  
Author(s):  
David A. Bushinsky ◽  
Walter R. Parker ◽  
Kristen M. Alexander ◽  
Nancy S. Krieger
Keyword(s):  
Metabolic Acidosis ◽  
Calcium Release ◽  
Calcium Absorption ◽  
Respiratory Acidosis ◽  
Urinary Calcium ◽  
Calcium Excretion ◽  
Calcium Efflux ◽  
Bicarbonate Concentration ◽  
Blood Ph ◽  
Urine Calcium Excretion

First published July 12, 2001; 10.1152/ajprenal.00355.2001.— A decrease in blood pH may be due to either a reduction in bicarbonate concentration ([HCO[Formula: see text]]; metabolic acidosis) or to an increase in Pco 2 (respiratory acidosis). In mammals, metabolic, but not respiratory, acidosis increases urine calcium excretion without altering intestinal calcium absorption, indicating that the additional urinary calcium is derived from bone. In cultured bone, chronic metabolic, but not respiratory, acidosis increases net calcium efflux ( J Ca), decreases osteoblastic collagen synthesis, and increases osteoclastic bone resorption. Metabolic acidosis increases bone PGE2production, which is correlated with J Ca, and inhibition of PGE2 production inhibits this acid-induced J Ca. Given the marked differences in the osseous response to metabolic and respiratory acidosis, we hypothesized that incubation of neonatal mouse calvariae in medium simulating respiratory acidosis would not increase medium PGE2 levels, as observed during metabolic acidosis. To test this hypothesis, we determined medium PGE2 levels and J Ca from calvariae incubated at pH ∼7.1 to model either metabolic (Met; [HCO[Formula: see text]] ∼11 mM) or respiratory (Resp; Pco 2 ∼83 Torr) acidosis, or at pH ∼7.5 as a control (Ntl). We found that after 24–48 and 48–51 h in culture, periods when cell-mediated J Capredominates, medium PGE2 levels and J Ca were increased with Met, but not Resp, compared with Ntl, and there was a direct correlation between medium PGE2 levels and J Ca. Thus metabolic, but not respiratory, acidosis induces the release of bone PGE2, which mediates J Ca from bone.

Mechanism of aluminum-induced calcium efflux from cultured neonatal mouse calvariae

1990 ◽  
Vol 258 (3) ◽  
pp. F583-F588 ◽  
Author(s):  
S. M. Sprague ◽  
D. A. Bushinsky
Keyword(s):  
Bone Mineral ◽  
Calcium Influx ◽  
Neonatal Mouse ◽  
Mineral Dissolution ◽  
Calcium Flux ◽  
Calcium Efflux ◽  
Dead Bone ◽  
45Ca Efflux ◽  
Dose Dependent

Aluminum has been shown to increase unidirectional 45Ca efflux from prelabeled bones in vitro; whether aluminum affects net calcium efflux and, if so, by what mechanism has not been studied. To examine the effects of aluminum on net calcium flux from bone we cultured live and dead neonatal mouse calvariae with and without graded concentrations of aluminum (10(-8) to 10(-5) M). Aluminum induced a dose-dependent net calcium efflux from live bone after 24 h, but not 3 h, which was similar in magnitude to that produced by 10(-8) M parathyroid hormone. The normal calcium influx into dead bone was not altered by aluminum. Release of beta-glucuronidase, a lysosomal enzyme released by osteoclasts, increased after a 24-h incubation in aluminum-containing medium and was correlated with net calcium efflux. Calcitonin, an inhibitor of osteoclastic bone mineral dissolution, abolished the increase in beta-glucuronidase release and nullified the aluminum-induced net calcium efflux. Thus aluminum induces cell-mediated net calcium efflux from bone and increases beta-glucuronidase release. Calcitonin inhibits the increase in both calcium efflux and beta-glucuronidase release, suggesting that aluminum stimulates osteoclasts to release bone mineral.

scholarly journals The effect of low initial medium pH on in vitro white poplar growth

2013 ◽  
pp. 67-80
Author(s):  
Branislav Kovacevic ◽  
Dragana Miladinovic ◽  
Marina Katanic ◽  
Zoran Tomovic ◽  
Sasa Pekec
Keyword(s):  
Biomass Accumulation ◽  
Dry Mass ◽  
White Poplar ◽  
Medium Ph ◽  
Shoot Height ◽  
Initial Ph ◽  
The Media ◽  
Initial Medium ◽  
Shoot Mass

The effect of low initial medium pH on shoot and root development of five white poplar (Populus alba L.) genotypes was tested. The shoot height, fresh mass of shoots per jar, dry mass of shoots per jar, number of roots, as well as the length of the longest root were measured and final pH of the media determined, after 35 days of culture in vitro. Three initial pH values of the medium were tested: 3.0, 4.0 and 5.5 as control. Agar solidification at pH 3.0 was not achieved after sterilization in autoclave, but it was successful after sterilizing in a microwave oven. The obtained results indicate that the tested genotypes are able to significantly influence the changes of media pH during culture. The effect of differences among the examined media was significant for biomass accumulation and final media pH. Generally, significantly higher values of fresh and dry shoot mass, shoot height and the longest root length were recorded on a medium with initial pH 3.0 then on a standard medium with pH 5.5.The implications of the obtained results for the improvement of in vitro propagation of white poplars are discussed.

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