İnvestigation of the effects of antioxidant applications during liposuction on the viability of adipose tissue-derived stem cells

Project no. 26327 · Status: Kapanmış (Yayın bekleniyor) · 2022

Summary

<p>Autologous&nbsp;fat&nbsp;grafting&nbsp;(lipofilling)&nbsp;is&nbsp;a&nbsp;widely&nbsp;used&nbsp;technique&nbsp;in&nbsp;Plastic&nbsp;and&nbsp;Reconstructive&nbsp;Surgery&nbsp;for&nbsp;the&nbsp;reconstruction&nbsp;of&nbsp;soft&nbsp;tissue&nbsp;defects&nbsp;and&nbsp;soft&nbsp;tissue&nbsp;augmentation.&nbsp;However,&nbsp;the&nbsp;most&nbsp;important&nbsp;limitation&nbsp;of&nbsp;this&nbsp;technique&nbsp;is&nbsp;the&nbsp;unpredictable&nbsp;nature&nbsp;of&nbsp;graft&nbsp;resorption&nbsp;and&nbsp;the&nbsp;occurrence&nbsp;of&nbsp;high-volume&nbsp;loss.&nbsp;The&nbsp;success&nbsp;of&nbsp;lipofilling&nbsp;depends&nbsp;on&nbsp;preserving&nbsp;the&nbsp;viability&nbsp;of&nbsp;adipose&nbsp;tissue-derived&nbsp;mesenchymal&nbsp;stem&nbsp;cells&nbsp;(AD-MSCs)&nbsp;within&nbsp;the&nbsp;stromal&nbsp;vascular&nbsp;fraction,&nbsp;preventing&nbsp;their&nbsp;loss,&nbsp;and&nbsp;initiating&nbsp;early&nbsp;angiogenesis.&nbsp;Mechanical&nbsp;trauma&nbsp;and&nbsp;ischemia&nbsp;occurring&nbsp;during&nbsp;liposuction&nbsp;may&nbsp;increase&nbsp;intracellular&nbsp;reactive&nbsp;oxygen&nbsp;species,&nbsp;leading&nbsp;to&nbsp;oxidative&nbsp;stress,&nbsp;which&nbsp;reduces&nbsp;stem&nbsp;cell&nbsp;viability&nbsp;and&nbsp;proliferation&nbsp;and&nbsp;consequently&nbsp;contributes&nbsp;to&nbsp;graft&nbsp;resorption.&nbsp;Mechanical&nbsp;trauma&nbsp;and&nbsp;ischemia&nbsp;during&nbsp;liposuction&nbsp;can&nbsp;lead&nbsp;to&nbsp;an&nbsp;increase&nbsp;in&nbsp;intracellular&nbsp;reactive&nbsp;oxygen&nbsp;species,&nbsp;causing&nbsp;oxidative&nbsp;stress.&nbsp;This&nbsp;oxidative&nbsp;stress&nbsp;can&nbsp;reduce&nbsp;stem&nbsp;cell&nbsp;viability&nbsp;and&nbsp;proliferation,&nbsp;ultimately&nbsp;leading&nbsp;to&nbsp;graft&nbsp;resorption.&nbsp;Therefore,&nbsp;this&nbsp;study&nbsp;was&nbsp;designed&nbsp;based&nbsp;on&nbsp;the&nbsp;hypothesis&nbsp;that&nbsp;the&nbsp;addition&nbsp;of&nbsp;pharmacological&nbsp;agents&nbsp;that&nbsp;reduce&nbsp;oxidative&nbsp;stress,&nbsp;such&nbsp;as&nbsp;N-acetylcysteine&nbsp;(NAC)&nbsp;and&nbsp;deferoxamine&nbsp;(DFO),&nbsp;to&nbsp;the&nbsp;liposuction&nbsp;solution&nbsp;could&nbsp;improve&nbsp;the&nbsp;efficiency&nbsp;of&nbsp;AD-MSC&nbsp;isolation.&nbsp;AD-MSCs&nbsp;obtained&nbsp;from&nbsp;lipoaspirates&nbsp;harvested&nbsp;using&nbsp;antioxidant-supplemented&nbsp;liposuction&nbsp;solutions&nbsp;were&nbsp;evaluated&nbsp;in&nbsp;terms&nbsp;of&nbsp;viability,&nbsp;proliferative&nbsp;capacity,&nbsp;and&nbsp;differentiation&nbsp;potential.&nbsp;Cells&nbsp;isolated&nbsp;from&nbsp;lipoaspirate&nbsp;samples&nbsp;obtained&nbsp;from&nbsp;healthy&nbsp;volunteer&nbsp;donors&nbsp;(n=6)&nbsp;were&nbsp;cultured&nbsp;as&nbsp;Control,&nbsp;NAC,&nbsp;and&nbsp;DFO&nbsp;groups.&nbsp;Cell&nbsp;viability,&nbsp;proliferative&nbsp;capacity,&nbsp;doubling&nbsp;time&nbsp;(DT),&nbsp;immunophenotypic&nbsp;characteristics,&nbsp;and&nbsp;differentiation&nbsp;potential&nbsp;were&nbsp;evaluated&nbsp;to&nbsp;determine&nbsp;the&nbsp;effects&nbsp;of&nbsp;preconditioning&nbsp;with&nbsp;NAC&nbsp;or&nbsp;DFO&nbsp;on&nbsp;AD-MSCs&nbsp;obtained&nbsp;from&nbsp;the&nbsp;same&nbsp;donor.&nbsp;The&nbsp;doubling&nbsp;time&nbsp;was&nbsp;significantly&nbsp;shorter&nbsp;in&nbsp;the&nbsp;DFO-&nbsp;and&nbsp;NAC-preconditioned&nbsp;groups&nbsp;in&nbsp;comparison&nbsp;to&nbsp;the&nbsp;control&nbsp;group&nbsp;(at&nbsp;24th&nbsp;hour:&nbsp;DFO-DT=23.10&nbsp;±&nbsp;0.14&nbsp;h&nbsp;and&nbsp;NAC-DT=21.08&nbsp;±&nbsp;0.37&nbsp;h&nbsp;vs.&nbsp;control-DT=28.17&nbsp;±&nbsp;0.25&nbsp;h,),&nbsp;and&nbsp;this&nbsp;difference&nbsp;was&nbsp;maintained&nbsp;at&nbsp;48th&nbsp;hour&nbsp;(DFO-DT=25.16&nbsp;±&nbsp;0.23&nbsp;h;&nbsp;NAC-DT=24.04&nbsp;±&nbsp;0.12&nbsp;h;&nbsp;vs&nbsp;control-DT=&nbsp;28.23&nbsp;±&nbsp;0.07&nbsp;h),&nbsp;and&nbsp;this&nbsp;reduction&nbsp;in&nbsp;DT&nbsp;indicated&nbsp;enhanced&nbsp;proliferative&nbsp;activity&nbsp;(p&nbsp;&lt;&nbsp;0.05).&nbsp;In&nbsp;MTT&nbsp;analysis,&nbsp;a&nbsp;significantly&nbsp;increased&nbsp;metabolic&nbsp;activity&nbsp;was&nbsp;observed&nbsp;in&nbsp;the&nbsp;NAC&nbsp;group&nbsp;at&nbsp;24th&nbsp;hour&nbsp;in&nbsp;comparison&nbsp;to&nbsp;both&nbsp;the&nbsp;control&nbsp;group&nbsp;and&nbsp;the&nbsp;DFO&nbsp;group&nbsp;(p&lt;0.001&nbsp;and&nbsp;p&lt;0.01,&nbsp;respectively).&nbsp;At&nbsp;48th&nbsp;hour,&nbsp;metabolic&nbsp;activity&nbsp;in&nbsp;the&nbsp;NAC&nbsp;group&nbsp;remained&nbsp;significantly&nbsp;elevated&nbsp;compared&nbsp;with&nbsp;the&nbsp;control&nbsp;group&nbsp;(p&lt;0.05).&nbsp;In&nbsp;contrast,&nbsp;the&nbsp;DFO&nbsp;group&nbsp;demonstrated&nbsp;only&nbsp;a&nbsp;slight&nbsp;increase&nbsp;in&nbsp;comparison&nbsp;to&nbsp;the&nbsp;control&nbsp;group,&nbsp;which&nbsp;did&nbsp;not&nbsp;reach&nbsp;statistical&nbsp;significance&nbsp;(p=NS).&nbsp;Flow&nbsp;cytometry&nbsp;analysis&nbsp;showed&nbsp;high&nbsp;positivity&nbsp;(95–100%)&nbsp;for&nbsp;the&nbsp;MSC&nbsp;biomarkers&nbsp;CD73&nbsp;and&nbsp;CD90&nbsp;in&nbsp;all&nbsp;groups,&nbsp;while&nbsp;CD45&nbsp;expression&nbsp;was&nbsp;negligible&nbsp;(0–3%),&nbsp;i.e.,&nbsp;negative.&nbsp;These&nbsp;results&nbsp;demonstrate&nbsp;that&nbsp;AD-MSCs&nbsp;obtained&nbsp;from&nbsp;lipoaspirate&nbsp;samples&nbsp;maintain&nbsp;their&nbsp;MSC&nbsp;phenotypes&nbsp;despite&nbsp;the&nbsp;application&nbsp;of&nbsp;drugs&nbsp;such&nbsp;as&nbsp;antioxidant&nbsp;NAC&nbsp;or&nbsp;hypoximimetic/Fe&nbsp;chelator&nbsp;DFO.&nbsp;Adipogenic,&nbsp;chondrogenic,&nbsp;and&nbsp;osteogenic&nbsp;differentiation&nbsp;capacities&nbsp;were&nbsp;preserved&nbsp;in&nbsp;all&nbsp;groups,&nbsp;while&nbsp;AD-MSCs&nbsp;obtained&nbsp;from&nbsp;NAC-&nbsp;or&nbsp;DFO-preconditioned&nbsp;lipoaspirates&nbsp;demonstrated&nbsp;an&nbsp;increased&nbsp;proliferative&nbsp;tendency.&nbsp;Morphological&nbsp;examination&nbsp;revealed&nbsp;that&nbsp;cell&nbsp;density&nbsp;and&nbsp;cellular&nbsp;organization&nbsp;were&nbsp;more&nbsp;pronounced&nbsp;in&nbsp;the&nbsp;NAC&nbsp;group,&nbsp;with&nbsp;cells&nbsp;exhibiting&nbsp;a&nbsp;compact&nbsp;and&nbsp;well-organized&nbsp;arrangement.</p><p>In&nbsp;conclusion,&nbsp;this&nbsp;clinical&nbsp;study&nbsp;demonstrated&nbsp;for&nbsp;the&nbsp;first&nbsp;time&nbsp;that&nbsp;adding&nbsp;NAC&nbsp;or&nbsp;DFO&nbsp;to&nbsp;the&nbsp;liposuction&nbsp;solution&nbsp;could&nbsp;increase&nbsp;the&nbsp;proliferation&nbsp;and&nbsp;biological&nbsp;activity&nbsp;of&nbsp;AD-MSCs&nbsp;obtained&nbsp;from&nbsp;lipoaspirates&nbsp;without&nbsp;altering&nbsp;or&nbsp;disrupting&nbsp;their&nbsp;MSC&nbsp;phenotype.&nbsp;These&nbsp;findings&nbsp;suggest&nbsp;that&nbsp;pharmacological&nbsp;preconditioning&nbsp;with&nbsp;antioxidant&nbsp;agents&nbsp;during&nbsp;liposuction&nbsp;may&nbsp;improve&nbsp;fat&nbsp;graft&nbsp;survival&nbsp;after&nbsp;lipofilling&nbsp;by&nbsp;reducing&nbsp;stem&nbsp;cell&nbsp;loss&nbsp;within&nbsp;the&nbsp;lipoaspirate,&nbsp;enhancing&nbsp;AD-MSC&nbsp;proliferation,&nbsp;and&nbsp;promoting&nbsp;differentiation&nbsp;into&nbsp;the&nbsp;desired&nbsp;cell&nbsp;types.&nbsp;Further&nbsp;studies&nbsp;in&nbsp;this&nbsp;area&nbsp;have&nbsp;the&nbsp;potential&nbsp;to&nbsp;enhance&nbsp;the&nbsp;effectiveness&nbsp;of&nbsp;regenerative/reparative&nbsp;medicine&nbsp;treatments,&nbsp;and&nbsp;our&nbsp;results&nbsp;will&nbsp;shed&nbsp;light&nbsp;on&nbsp;further&nbsp;research&nbsp;in&nbsp;this&nbsp;field.&nbsp;</p><p></p>

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