Erodible thermogelling hydrogels for localized mitochondrial transplantation to the spinal cord.
Erodible thermogelling hydrogels for localized mitochondrial transplantation to the spinal cord.
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可侵蚀的热凝水水凝胶，用于局部线粒体移植到脊髓。
DOI：
10.1016/j.mito.2022.04.002
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发表时间：
2022-05
期刊：
影响因子：
4.4
通讯作者：
Rabchevsky, Alexander G.
中科院分区：
文献类型：
--
作者：
Patel, Samir P.;Michael, Felicia M.;Khan, M. Arif;Duggan, Brian;Wyse, Sam;Darby, Daniel R.;Chaudhuri, Krishnaroop;Pham, Jonathan T.;Gollihue, Jenna;DeRouchey, Jason E.;Sullivan, Patrick G.;Dziubla, Tom D.;Rabchevsky, Alexander G.
我们开发了一种热凝胶、可侵蚀的水凝胶系统，用于体内活性线粒体的局部传递，并用特定的染料标记移植的线粒体和/或用红色荧光蛋白(RFP)标记的转基因线粒体。我们还利用细胞系优化了一种由甲基纤维素和透明质酸组成的水凝胶，旨在保护生物能量，同时促进线粒体的释放。我们进一步研究了将同种或异种线粒体移植到各自的细胞系中如何影响宿主细胞的代谢，通过MTS试验进行了测量。我们发现，在37℃下，水凝胶在20min内释放了70%的线粒体，60min后水凝胶释放的线粒体的呼吸能力比没有凝胶的线粒体大，线粒体的MTR标记并不是不可磨灭的。从改良的SH-SY5Y人神经母细胞瘤细胞中分离的RFP标记的转基因线粒体对单纯SH-SY5Y细胞和大鼠PC-12细胞都显示出有效的摄取，尤其是当从水凝胶中释放时。该水凝胶既能在体外生理条件下保护线粒体，又能在60min内局部固化和扩散。为了评估代谢效应，将不同浓度的SH-SY5Y或PC-12细胞来源的线粒体移植到两种细胞系中，均导致移植后6小时和24小时的代谢显著增加。或者，移植大鼠大脑和脊髓组织中最高浓度的线粒体会在24小时后降低代谢活动。随着水凝胶的改进，我们正在进一步研究这种代谢变化是由于细胞增殖的变化还是由于单个宿主细胞中并入的外源线粒体的数量的变化。
We developed a thermal-gelling, erodible hydrogel system for localized delivery of viable mitochondria in vivo, as well as labeled transplanted mitochondria with specific dyes and/or genetically modified mitochondria tagged with red fluorescence protein (RFP). We also employed cell lines to optimize a hydrogel composed of methylcellulose and hyaluronic acid designed to preserve bioenergetics while facilitating mitochondrial release. We further investigated how transplantation of allogeneic or xenogeneic mitochondria into respective cell lines affects host cellular metabolism, as measured by MTS assay. We found that 70% of mitochondria are released from the hydrogel within 20 min at 37 °C, that the respiratory capacity of hydrogel-released mitochondria over 60 min was greater than those without gel, and that MTR-labeling of mitochondria is not indelible. RFP-tagged transgenic mitochondria isolated from modified SH-SY5Y human neuroblastoma cells showed effective uptake into both naïve SH-SY5Y cells and rat PC-12 cells, notably when released from hydrogel. The hydrogel both protected the mitochondria at physiological conditions in vitro while solidifying and diffusing within 60 min locally in situ. To assess metabolic effects, both cell lines were transplanted with different concentrations of SH-SY5Y or PC-12 cell line-derived mitochondria and all resulted in significant increases in metabolism at 6- and 24-hour after transplantation. Alternatively, transplanted mitochondria at highest concentration from rat brain and spinal cord tissues reduced metabolic activities after 24-hour. Along with hydrogel refinements, we are further investigating whether such metabolic changes are due to alterations in cell proliferation or the number of exogenous mitochondria incorporated into individual host cells.
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影响因子：
4.4
作者：
Patel, Samir P.;Gamboa, Jorge L.;McMullen, Colleen A.;Rabchevsky, Alexander;Andrade, Francisco H.
通讯作者：
Andrade, Francisco H.
影响因子：
4.2
作者：
Patel SP;Sullivan PG;Pandya JD;Rabchevsky AG
通讯作者：
Rabchevsky AG
影响因子：
4.4
作者：
McCully, James D.;del Nido, Pedro J.;Emani, Sitaram M.
通讯作者：
Emani, Sitaram M.
DOI：
10.1152/ajpheart.00567.2008
发表时间：
2009-01-01
影响因子：
4.8
作者：
McCully, James D.;Cowan, Douglas B.;Levitsky, Sidney
通讯作者：
Levitsky, Sidney
影响因子：
3
作者：
Gollihue, Jenna L.;Patel, Samir P.;Rabchevsky, Alexander G.
通讯作者：
Rabchevsky, Alexander G.