The effect of ultrasound stimulation on the gene and protein expression of chondrocytes seeded in chitosan scaffolds


Hasanova G. I., Noriega S. E., Mamedov T., Thakurta S. G., Turner J. A., Subramanian A.

JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, cilt.5, sa.10, ss.815-822, 2011 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 5 Sayı: 10
  • Basım Tarihi: 2011
  • Doi Numarası: 10.1002/term.384
  • Dergi Adı: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.815-822
  • Anahtar Kelimeler: chondrocytes, chitosan scaffolds, ultrasound stimulation, gene expression, LOW-INTENSITY ULTRASOUND, INTERMITTENT PHYSIOLOGICAL PRESSURE, MESENCHYMAL STEM-CELLS, GROWTH-FACTOR-BETA, PULSED ULTRASOUND, MATRIX PRODUCTION, ARTICULAR-CARTILAGE, REGENERATING CARTILAGE, HYDROSTATIC-PRESSURE, DYNAMIC COMPRESSION
  • Akdeniz Üniversitesi Adresli: Hayır

Özet

Both pulsed-and square-wave, low-intensity ultrasound (US) signals have been reported to impact chondrocyte function and biosynthetic activity. In this study, a low-intensity diffuse ultrasound (LIDUS) signal at 5.0 MHz (0.14 mW/cm(2)) was employed to stimulate bovine chondrocytes seeded in three-dimensional (3D) chitosan-based matrices. While the duration of application was constant at 51 s, US was applied once, twice, four times and eight times/day, and the impacts of US on the biosynthetic activity of chondrocytes and the expression of chondrocyte-specific genes were evaluated. When stimulated with continuous US for predetermined time intervals, chondrocytes had higher levels of type II collagen, aggrecan, L-Sox5 and Sox9 mRNA expression when compared to controls; however, under the same conditions, the expression of MMP-3 was downregulated. Interestingly, both Sox5 and Sox9 genes coordinately responded to changes in US stimulation and generally mirrored the response of collagen type II transcript to changes in US stimulation. RT-PCR analysis revealed that US stimulation increased the gene expression of cell-surface integrins alpha 5 and beta 1. The expression of integrins alpha 2 was downregulated by US treatment, suggesting that multiple integrin subunits may be involved in the regulation of chondrocytic function in response to US stimuli. The enhancement in the abundance of the mRNA transcripts upon US stimulation was observed to correlate with the protein expression of collagen type I, collagen type II, and integrins alpha 5 and beta 1. In conclusion, the US stimulation regimen employed was shown to modulate the proliferative capacity, biosynthetic activity and integrin mRNA expression of articular chondrocytes maintained in 3D matrices. Copyright (C) 2011 John Wiley & Sons, Ltd.