The effects of cold storage and endotoxin challenge on osteoblast viability and interleukin-6 production

Frederick C. Bisch, Benjamin S. Hanson, Norma H. Best, David W. Craft

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Autogenous hip marrow is an excellent source of pluripotential cells for regenerative procedures. However, before this treatment modality can be employed a method to attenuate osteoclast activity must be developed. The shock of cold storage (4°C) is thought to abate osteoclast activity through the downregulation of osteolytic cytokines produced by osteoblasts. The objective of this study was to evaluate the effects of cold storage (4°C) and endotoxin challenge on bone cell culture viability and interleukin-6 (IL-6) production. These cells (osteoblasts) were primarily harvested from murine calvaria utilizing sequential digestions, separated by density gradient and combined. Twelve-well cell culture plates were inoculated with 2×104 cells/ml and placed in cold storage for 1-14 d. After cold storage the cultures were then incubated at 37°C for 1-20 d. A set of replicate plates was also challenged with 10 ng/ml endotoxin upon incubation at 37°C for 4 consecutive days. Cells were evaluated daily for alkaline phosphatase activity. Cell culture supernatants were also collected daily and batch assayed for IL-6 production. Cell cultures did not survive more than 48 h of cold storage. There was a decrease in IL-6 secretion in all refrigerated cultures and a significant decrease in those cells refrigerated for 48 h versus control cultures (p<0.05). Replicate cultures treated with endotoxin secreted significantly increased amounts of IL-6 in both the control cultures and the cultures exposed to 24 h of cold storage versus non-endotoxin-treated control cultures (p<0.05). These observations suggest that after 48 h of cold storage autogenous marrow may be safe to use because of the dramatic decrease in IL-6 production by osteoblasts.

Original languageEnglish (US)
Pages (from-to)150-155
Number of pages6
JournalJournal of Periodontal Research
Volume33
Issue number3
StatePublished - Apr 1 1998

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Osteoblasts
Endotoxins
Interleukin-6
Cell Culture Techniques
Osteoclasts
Bone Marrow
Skull
Alkaline Phosphatase
Hip
Digestion
Shock
Cell Survival
Down-Regulation
Cytokines
Bone and Bones

All Science Journal Classification (ASJC) codes

  • Periodontics

Cite this

Bisch, Frederick C. ; Hanson, Benjamin S. ; Best, Norma H. ; Craft, David W. / The effects of cold storage and endotoxin challenge on osteoblast viability and interleukin-6 production. In: Journal of Periodontal Research. 1998 ; Vol. 33, No. 3. pp. 150-155.
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The effects of cold storage and endotoxin challenge on osteoblast viability and interleukin-6 production. / Bisch, Frederick C.; Hanson, Benjamin S.; Best, Norma H.; Craft, David W.

In: Journal of Periodontal Research, Vol. 33, No. 3, 01.04.1998, p. 150-155.

Research output: Contribution to journalArticle

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