Chondrocytes in cartilage are embedded in a matrix containing a high concentration of proteoglycans and hence of fixed negative charges. Their extracellular ionic environment is thus different from that of most cells, with extracellular Na+ being 250-350 mM and extracellular osmolality 350-450 mOsm. When chondrocytes are isolated from the matrix and incubated in standard culture medium (DMEM; osmolality 250-280 mOsm), their extracellular environment changes sharply. We incubated isolated bovine articular chondrocytes and cartilage slices in DMEM whose osmolality was altered over the range 250-450 mOsm by Na+ or sucrose addition. 35S-sulphate and 3H-proline incorporation rates were at a maximum when the extracellular osmolality was 350-400 mOsm for both freshly isolated chondrocytes and for chondrocytes in cartilage. The incorporation rate per cell of isolated chondrocytes was only 10% that of chondrocytes in situ both 4 and 24 hours after isolation. For freshly isolated chondrocytes, the rate increased 30-50% in DMEM to which NaCl or sucrose had been added to increase osmolality. In chondrocytes incubated overnight in DMEM, the rate was greatest in DMEM of normal osmolality and fell from the maximum in proportion to the change in osmolality. The effects of sucrose addition on incorporation rates were similar but not identical to those of Na+ addition. Changes in cell volume might be linked to changes in synthesis rates since the cell volume of chondrocytes (measured by Coulter-counter) increased 30-40% when the cells were removed from their in situ environment into DMEM. Synthesis rates can thus be partly regulated by changes in extracellular osmolality, which in cartilage is controlled by proteoglycan concentration. This provides a mechanism by which the chondrocytes can rapidly respond to changes in extracellular matrix composition.

The direct effects of hydrostatic pressure on matrix synthesis in articular cartilage can be studied independently of the other factors that change during loading. We have found that the influence of hydrostatic pressure on incorporation rates of 35SO4 and [3H]proline into adult bovine articular cartilage slices in vitro depends on the pressure level and on the time at pressure. Pressures in the "physiological" range (5-15 MPa) applied for 20 s or for 5 min could stimulate tracer incorporation (30-130%) during the following 2 h, but higher pressures (20-50 MPa) had no effect on incorporation rates. The degree of stimulation in cartilage obtained from different animals was found to vary; in some animals none was seen. Stimulation also varied with position along the joint. Physiological pressures (5-10 MPa) applied continuously for the 2-h incubation period also stimulated incorporation rates, but pressures greater than 20 MPa always produced a decrease that was related to the applied pressure and that was reversible. These results suggests that the hydrostatic pressure that occurs during loading is a signal that can stimulate matrix synthesis rates in articular cartilage.

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The overall internal pH of the acid-tolerant green alga, Chlorella saccharophila, was determined in the light and in the dark by the distribution of 5,5-dimethyl-2-14Cloxazolidine-2,4-dione (1[4qDMO) or ['Cjbenzoic acid (['4CIBA) between the cells and the surrounding medium.['CIDMO was used at external pH of 5.0 to 7.5 while ['4C]BA was used in the range pH 3.0 to pH 5.5. Neither compound was metabolized by the algal cells and intracellular binding was minimal. The internal pH of the algae obtained with the two compounds at external pH values of 5.0 and 5.5 were in good agreement. The internal pH of C. saccharophila remained relatively constant at pH 7.3 over the external pH range of pH 5.0 to 7.5. Below pH 5.0, however, there was a gradual decrease in the internal pH to 6.4 at an external pH of 3.0. The maintenance of a constant internal pH requires energy and the downward drift of internal pH with a drop in external pH may be a mechanism to conserve energy and allow growth at acid pH.Microscopic algae generally have pH optima for growth and photosynthesis in the neutral to alkaline pH range. There are some species, however, which can grow and photosynthesize in acid conditions and studies with procaryotes have shown that it is necessary for such acidophilic organisms to maintain a neutral internal pH (17). Our knowledge of the variation in internal pH of photosynthetic microorganisms with changes in external pH is limited to only a few species of green algae and cyanobacteria (7, 12, 13,16).The internal pH of microorganisms can be conveniently and accurately measured by determining the distribution of a radioactively-labeled weak acid or weak base between the intracellular space and an external solution of known pH. It is assumed in this method that the uncharged labeled compound passively diffuses into the cell so that at equilibrium the concentrations of the uncharged species inside and outside the cell will be equal. Thus, from the pK of the labeled compound, the external pH, and the measured concentrations of the compound inside and outside the cell, the internal pH can be calculated (5). To accurately measure the equilibrium concentrations of the compound in the cells and the medium, the pH of the medium should be ± 1.0 to 1.

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Various physiological characteristics of photosynthesis in the unicellular red alga Porphyridium cruentum Naegeli have been investigated. The rate of photosynthesis was optimal at 25° C and pH 7.5 and was not inhibited by 21% oxygen over a temperature range of 5 to 35° C. Kinetics of whole cell photosynthesis as a function of substrate concentration gave a K1/2, (CO2) of 0.3 μM. CO2 compensation point, measured in a closed system at pH 7.5, was a constant 6.7 m̈L · L−1 over the temperature range 15 to 30° C and was unaffected by O2 concentration. Whole cell photosynthesis, measured in a closed system at alkaline pH, showed that the rates of oxygen evolution were greatly in excess of the rate of CO2 supply from the spontaneous dehydration of HCO3− in the medium. This indicates that bicarbonate is utilized by the cell to support this photosynthetic rate. These physiological characteristics of Porphyridium cruentum are consistent with the hypothesis that this alga transports bicarbonate across the plasmalemma.