低強度脈衝超音波及高葡萄糖對骨細胞之效應

Effect of Low-Intensity Pulsed Ultrasound and High Glucose on Bone Cells

陳琇慧(Hsiu-Hui Chen)

中原大學/工學院/生物醫學工程研究所/博士(2013年)

https://doi.org/10.6840/CYCU.2013.00393

摘要

糖尿病患易有高骨質疏鬆症及骨折風險，但糖尿病與骨生成、骨吸收等關係仍有爭議。治療性超音波已被證實可促進骨折癒合，且可藉由調整參數促進或抑制細胞之生長，然而，不同超音波刺激參數對骨細胞功能的影響仍不清楚。因此，為了解低強度脈衝超音波(low-intensity pulsed ultrasound, LIPUS)對骨細胞在正常及高葡糖之生物效應，本研究量測換能器聲場特性、探討不同LIPUS參數對骨母細胞的影響、LIPUS及高葡萄對骨母細胞生長的影響、以及LIPUS對巨噬細胞分化蝕骨細胞的影響。實驗結果顯示：LIPUS可促進骨母細胞存活率、alkaline phosphatase (ALP)活性、骨礦化、ALP與osteocalcin (OCN) mRNA表現，且具有劑量依賴性。而高葡萄糖則會抑制細胞存活率、adenosine triphosphate (ATP)表現、細胞爬行、OCN mRNA表現；增加ALP、osteoprotegerin、heat shock protein 90 (Hsp90) mRNA表現。在高葡萄糖濃度下，LIPUS可促進細胞存活率、ATP表現、細胞爬行、Hsp90與OCN mRNA表現。此外，LIPUS可抑制蝕骨細胞tartrate-resistant acid phosphatase (TRAP)活性、骨吸收作用，以及TRAP、receptor-activated nuclear factor-kappa B、cathepsine K、matrix metalloproteinase-9 mRNA表現。綜合以上結果證實：適當LIPUS刺激參數可有效促進骨母細胞之生長與分化，且可降低高葡萄糖下骨母細胞生長以及細胞爬行被抑制之效應，並可抑制蝕骨細胞骨分化，此結果有助於LIPUS刺激參數之選擇、提供超音波生物效應對骨細胞在骨再生之研究觀點，相關的細胞生物物理機制仍然需要進一步探討，期望未來可為糖尿病與骨相關疾病提供創新的治療方式。

關鍵字

骨母細胞 ； 糖尿病 ； 高葡萄糖 ； 低強度脈衝超音波 ； 蝕骨細胞

並列摘要

Diabetes mellitus (DM) is a metabolic disorder that predisposes patients to a high risk of skeletal disorders such as osteoporosis and bone fracture. However, the relationships among DM, bone formation, and bone resorption remain controversial. The promotion or inhibition of cell growth has been shown to be regulated by therapeutic ultrasound in a parameter-dependent manner. Nevertheless, the cellular functions affected by various ultrasound insonifications are not well understood. Therefore, to determine the effects of low-intensity pulsed ultrasound (LIPUS) and high glucose (HG) on bone cells, we investigated the characteristics of transducer and sound fields; the effects of various LIPUS insonifications on the growth of osteoblasts; and the effect of LIPUS insonification on the growth of osteoblasts under normal and HG conditions. We also examined the effect of LIPUS on osteoclastogenesis in RAW264.7 cells.
The results showed that LIPUS insonification enhanced cell viability, alkaline phosphatase (ALP) activity, ALP and osteocalcin (OCN) mRNA expression, and mineralization of osteoblasts in a dose-dependent manner. Cell viability, adenosine triphosphate (ATP), cell migration, and OCN mRNA levels were lower in the high-glucose medium whereas ALP mRNA, osteoprotegerin, and heat shock protein 90 (Hsp90) levels were higher. On the other hand, LIPUS insonification increased cell viability, ATP levels, cell migration, Hsp90 and OCN mRNA levels of osteoblasts in the HG medium. Besides, the insonified osteoclasts had lower levels of tartrate-resistant acid phosphatase (TRAP) activity; pit formation; TRAP, Cathepsin K, receptor-activated nuclear factor-kappa B, and matrix metalloproteinase-9 mRNA. In conclusion, an adequate insonification enhances the growth and differentiation of osteoblasts. LIPUS decreased the inhibitory effect on osteoblasts associated with HG concentration and inhibited the differentiation of osteoclasts from RAW 264.7 cells. These findings may support therapeutic ultrasound, particularly the selection of appropriate parameters and understanding the ultrasonic biological effects of LIPUS on bone cells during bone generation. The biophysical mechanisms underlying the effects of ultrasound on bone cells need to be investigated further to facilitate the development of novel treatments for bone diseases associated with DM.

並列關鍵字

osteoblast ； diabetes mellitus ； high glucose ； low-intensity pulsed ultrasound ； osteoclast

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