Characterization of dense artificial connective tissues generated in a newly designed bioreactor

Hironobu Iwashiro, Satoshi Hosoya, Kazuya Hirai, Takehiko Mima, Shihoka Ohashi, Takashi Aihara, Shuhei Ito, Susumu Ohara, Eijiro Adachi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Dense connective tissues were generated simultaneously by accumulating collagen fibrils and fibroblasts on stainless steel mesh using a bioreactor system that we designed. The advantage of our system is that the artificial connective tissues can be generated within 24 hr in the absence of inhibitors against matrix metalloproteinases. The fibroblasts were suspended in 200 mL of Dulbecco's modified Eagle's medium containing 10% fetal bovine serum and 0.5 mg/mL type I collagen. The mixed solution was circulated in two types of bioreactors with cylindrical or vertical configurations to generate luminal or parenchymal tissues, respectively. The gelatin zymography showed that MMPs were first detected in the media after 8 hr from the start of circulation and reached the highest levels on day 3. Glossy white aggregates, 13 mm in thickness, depending on the circulation period, accumulated on mesh grids. Fibroblasts were embedded in the network of collagen fibrils and possessed oval nuclei with or without prominent cell processes to form a bipolar shape. We could not observe distended cisternae of the endoplasmic reticula, the Golgi apparatus, or exploded mitochondria, showing hypoxic degenerative alterations of fibroblasts in dense connective tissues. The artificial tissues generated by our system will be useful for biological studies and transplantation therapy.

Original languageEnglish
Pages (from-to)340-352
Number of pages13
JournalConnective Tissue Research
Volume52
Issue number4
DOIs
Publication statusPublished - Aug 1 2011
Externally publishedYes

Keywords

  • Bioreactor
  • Collagen fibrils
  • Dermis
  • Electron microscopy
  • Fibroblasts

ASJC Scopus subject areas

  • Rheumatology
  • Biochemistry
  • Orthopedics and Sports Medicine
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Iwashiro, H., Hosoya, S., Hirai, K., Mima, T., Ohashi, S., Aihara, T., Ito, S., Ohara, S., & Adachi, E. (2011). Characterization of dense artificial connective tissues generated in a newly designed bioreactor. Connective Tissue Research, 52(4), 340-352. https://doi.org/10.3109/03008207.2010.531801