Behavior of Hyaluronic ACID from Gingival Epithelium and Connective Tissue on the Analytical Ultracentrifuge

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Abstract
The molecular weights of hyaluronic acid (HA) isolated from separated specimens of human gingival epithelium and connective tissue as well as standard hyaluronic acid preparations have been estimated. The values were determined following substitution of sedimentation values into a previously determined empirical relationship between the reciprocal of the sedimentation coefficient at zero concentration (s−1)0 and molecular weights estimated by sedimentation-diffusion (MsD). The values of (s−1)0 for connective tissue and standard low-molecular weight HA preparations obtained by linear regression of all points indicated molecular weights (MsD) of 340,000 and 205,000 respectively. However, epithelial and standard high-molecular weight HA behaved differently during ultracentrifugation generating a curvilinear relationship between s−1 and concentration. Nevertheless linear extrapolation of a line of best fit of the very lowest concentrations (those which approached zero concentration) provided molecular weight estimates of 860,000 and 2,500,000 respectively. Moreover, similar treatment of s−1 values derived from the previously published data of Laurent, Ryan and Pietruszkiewicz1 has validated the use of linear regression of s−1 at the lower concentrations alone, to calculate high-molecular weight HA. The curvilinear relationship for s−1 throughout the whole concentration range (0.15–2.3 mg/ml) has been regarded only as a qualitative indication that the HA samples are of relatively high-molecular weight, while a straight line through such data points implies a qualitatively lower molecular weight for HA.