Fluorometric determination of DNA in cartilage of various species

Theodore R. Oegema, Barbara J. Carpenter, Roby C. Thompson

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

A sensitive, modified 3,5‐diaminobenzoic acid (DABA), fluorometric DNA assay was developed and compared to mithramycin and ethidium bromide assays in determining the DNA content of dense connective tissues including: Swarm rat chondrosarcoma, rabbit, dog, monkey, and most importantly, adult human articular cartilage. In the more cellular cartilages, the three methods gave equivalent results. However, in the relatively acellular human cartilage, the DABA method was shown to be superior. Both the mithramycin and ethidium bromide gave falsely high values compared to the DABA method, which by subtraction after DNase digestion approached the DABA value. The latter was completely DNase sensitive. With the DABA method, the DNA content of human cartilage can be obtained on less than 5 mg wet weight of fresh, alcohol‐fixed, or lyophilized material. While the DNA can also be released by digestion with papain or protease from Streptomyces griseus, proteinase K was preferable. The comparison of literature values for other fluorometric and spectrophotometric assays of human cartilage suggest these methods overestimate human articular cartilage DNA concentrations, whereas the DABA values were in line with those predicted from previous morphometric analysis. Thus, the modified method represents an improvement in DNA analysis of dense connective tissues.

Original languageEnglish (US)
Pages (from-to)345-351
Number of pages7
JournalJournal of Orthopaedic Research
Volume1
Issue number4
DOIs
StatePublished - Jan 1 1983

Fingerprint

Cartilage
DNA
Plicamycin
Ethidium
Deoxyribonucleases
Articular Cartilage
Connective Tissue
Digestion
Streptomyces griseus
Endopeptidase K
Chondrosarcoma
Papain
Haplorhini
DABA
Peptide Hydrolases
Dogs
Rabbits
Weights and Measures
Acids

Keywords

  • Cartilage
  • Connective tissue
  • DNA
  • Fluorometric analysis
  • Human

Cite this

Fluorometric determination of DNA in cartilage of various species. / Oegema, Theodore R.; Carpenter, Barbara J.; Thompson, Roby C.

In: Journal of Orthopaedic Research, Vol. 1, No. 4, 01.01.1983, p. 345-351.

Research output: Contribution to journalArticle

Oegema, Theodore R. ; Carpenter, Barbara J. ; Thompson, Roby C. / Fluorometric determination of DNA in cartilage of various species. In: Journal of Orthopaedic Research. 1983 ; Vol. 1, No. 4. pp. 345-351.
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