Preparation of Rat Gingival Mitochondria with an Improved Isolation Method

Table 2

Respiratory control ratio (RCR) and adenosine diphosphate/oxygen (ADP/O) ratio in mitochondrial fractions from rat gingival tissues.

Concentration of; collagenase [%(w/v)]	Respiratory control ratio	ADP/O ratio
Homogenized for 40 sec
0.120	1.27 ± 0.04	1.20 ± 0.06
0.125	1.54 ± 0.02	1.39 ± 0.05
0.130	1.43 ± 0.04	1.25 ± 0.02
Homogenized for 50 sec
0.115	1.45 ± 0.03	1.36 ± 0.03
0.120	1.80 ± 0.05∗,∗∗	1.65 ± 0.03∗,∗∗
0.125	1.39 ± 0.04	1.28 ± 0.03
Homogenized for 60 sec
0.115	1.50 ± 0.03	1.42 ± 0.02
0.120	1.57 ± 0.03	1.49 ± 0.01
0.125	1.33 ± 0.05	1.23 ± 0.03

The respiratory control ratio (RCR) and adenosine diphosphate/oxygen (ADP/O) ratio of the mitochondrial (Mt) fractions which were obtained by varying the concentration of collagenase and the time of homogenization were measured. After measuring states 1, 3, and 4, the respiratory activity of gingival Mt was assessed by measuring (1) the respiratory control ratio (RCR): the ratio of the respiratory rate (State 3) in the presence of ADP to the rate (State 4) following the expenditure of ADP and (2) adenosine diphosphate/oxygen (ADP/O) ratio: the ratio of ADP removed from the media to the amount of oxygen consumed, according to the Estabrook method [15].
∗Significantly different in terms of the highest values between 40 sec and 50 sec homogenizing at 𝑃<.05.
∗∗ Significantly different in terms of the highest values between 50 sec and 60 sec homogenizing at 𝑃<.05.