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首頁> 外文學(xué)位 >The Use of a 3D Perfusion Bioreactor with Osteoblasts and Osteoblast/Endothelial Cell Co-cultures to Improve Tissue-Engineered Bone.
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The Use of a 3D Perfusion Bioreactor with Osteoblasts and Osteoblast/Endothelial Cell Co-cultures to Improve Tissue-Engineered Bone.

機譯：將3D灌注生物反應(yīng)器與成骨細胞和成骨細胞/內(nèi)皮細胞共培養(yǎng)物一起使用，以改善組織工程骨。

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摘要

The delivery of oxygen, nutrients, and the removal of waste are essential for cellular survival. Culture systems for 3D bone tissue engineering have addressed this issue by utilizing perfusion flow bioreactors that stimulate osteogenic activity through the delivery of oxygen and nutrients by low-shear fluid flow. It is also well established that bone responds to mechanical stimulation, but may desensitize under continuous loading. While perfusion flow and mechanical stimulation are used to increase cellular survival in vitro, 3D tissue-engineered constructs face additional limitations upon in vivo implantation. As it requires significant amounts of time for vascular infiltration by the host, implants are subject to an increased risk of necrosis. One solution is to introduce tissue-engineered bone that has been pre-vascularized through the co-culture of osteoblasts and endothelial cells on 3D constructs.;It is unclear from previous studies: 1) how 3D bone tissue constructs will respond to partitioned mechanical stimulation, 2) how gene expression compares in 2D and in 3D, 3) how co-cultures will affect osteoblast activity, and 4) how perfusion flow will affect co-cultures of osteoblasts and endothelial cells. We have used an integrated approach to address these questions by utilizing mechanical stimulation, perfusion flow, and a co-culture technique to increase the success of 3D bone tissue engineering. We measured gene expression of several osteogenic and angiogenic genes in both 2D and 3D (static culture and mechanical stimulation), as well as in 3D cultures subjected to perfusion flow, mechanical stimulation and partitioned mechanical stimulation. Finally, we co-cultured osteoblasts and endothelial cells on 3D scaffolds and subjected them to long-term incubation in either static culture or under perfusion flow to determine changes in gene expression as well as histological measures of osteogenic and angiogenic activity.;We discovered that 2D and 3D osteoblast cultures react differently to shear stress, and that partitioning mechanical stimulation does not affect gene expression in our model. Furthermore, our results suggest that perfusion flow may rescue 3D tissue-engineered constructs from hypoxic-like conditions by reducing hypoxia-specific gene expression and increasing histological indices of both osteogenic and angiogenic activity. Future research to elucidate the mechanisms behind these results may contribute to a more mature bone-like structure that integrates more quickly into host tissue, increasing the potential of bone tissue engineering.
機譯：氧氣，營養(yǎng)物質(zhì)的輸送和廢物的清除對于細胞存活至關(guān)重要。用于3D骨組織工程的培養(yǎng)系統(tǒng)通過利用灌注流生物反應(yīng)器解決了這個問題，該反應(yīng)器通過低剪切液流通過輸送氧氣和營養(yǎng)來刺激成骨活性。眾所周知，骨骼對機械刺激有反應(yīng)，但在連續(xù)負荷下可能會脫敏。雖然使用灌注流和機械刺激來提高體外細胞存活率，但3D組織工程構(gòu)建的構(gòu)建體在體內(nèi)植入時面臨其他限制。由于宿主需要大量時間進行血管浸潤，因此植入物的壞死風(fēng)險增加。一種解決方案是將已通過成骨細胞和內(nèi)皮細胞的共培養(yǎng)在3D構(gòu)造上進行血管形成的組織工程化骨骼引入；從先前的研究中尚不清楚：1）3D骨組織構(gòu)造將如何響應(yīng)分區(qū)的機械刺激； 2）基因表達在2D和3D中的比較方式； 3）共培養(yǎng)將如何影響成骨細胞的活性； 4）灌注流量將如何影響成骨細胞和內(nèi)皮細胞的共培養(yǎng)。我們已經(jīng)使用一種綜合方法來解決這些問題，方法是利用機械刺激，灌注流和共培養(yǎng)技術(shù)來提高3D骨組織工程的成功率。我們在2D和3D（靜態(tài)培養(yǎng)和機械刺激）中以及在經(jīng)歷灌注流，機械刺激和分區(qū)機械刺激的3D培養(yǎng)物中，測量了幾種成骨和血管生成基因的基因表達。最后，我們在3D支架上共培養(yǎng)成骨細胞和內(nèi)皮細胞，并在靜態(tài)培養(yǎng)或灌注流下對其進行長期孵育，以確定基因表達的變化以及成骨和血管生成活性的組織學(xué)測量方法。 2D和3D成骨細胞培養(yǎng)物對剪切應(yīng)力的反應(yīng)不同，并且在我們的模型中分配機械刺激不會影響基因表達。此外，我們的結(jié)果表明，灌注流可通過減少缺氧特異性基因表達并增加成骨和血管生成活性的組織學(xué)指標，從缺氧樣條件中拯救3D組織工程構(gòu)造。闡明這些結(jié)果背后機制的未來研究可能有助于形成更成熟的骨樣結(jié)構(gòu)，這種結(jié)構(gòu)可以更快地整合到宿主組織中，從而增加了骨組織工程學(xué)的潛力。

著錄項

  • 作者

    Barron, Matthew J.;

  • 作者單位

    Michigan Technological University.;

  • 授予單位 Michigan Technological University.;
  • 學(xué)科 Engineering Biomedical.
  • 學(xué)位 Ph.D.
  • 年度 2010
  • 頁碼 116 p.
  • 總頁數(shù) 116
  • 原文格式 PDF
  • 正文語種 eng
  • 中圖分類
  • 關(guān)鍵詞

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