Mao Yong, John Nikita, Protzman Nicole M, Kuehn Adam, Long Desiree, Sivalenka Raja, Junka Radoslaw A, Gosiewska Anna, Hariri Robert J, Brigido Stephen A
Laboratory for Biomaterials Research, Department of Chemistry and Chemical Biology, Rutgers University, 145 Bevier Rd., Piscataway, NJ, 08854, USA.
Healthcare Analytics, LLC, 78 Morningside Dr., Easton, PA, 18045, USA.
J Exp Orthop. 2022 Jul 18;9(1):69. doi: 10.1186/s40634-022-00509-4.
Injectable connective tissue matrices (CTMs) may promote tendon healing, given their minimally invasive properties, structural and biochemical extracellular matrix components, and capacity to fill irregular spaces. The purpose of this study is to evaluate the effects of placental CTMs on the cellular activities of human tenocytes. Decellularization, the removal of cells, cell fragments, and DNA from CTMs, has been shown to reduce the host's inflammatory response. Therefore, the authors hypothesize that a decellularized CTM will provide a more cell-friendly matrix to support tenocyte functions.
Three human placental CTMs were selected for comparison: AmnioFill® (A-CTM), a minimally manipulated, non-viable cellular particulate, BioRenew™ (B-CTM), a liquid matrix, and Interfyl® (I-CTM), a decellularized flowable particulate. Adhesion and proliferation were evaluated using cell viability assays and tenocyte migration using a transwell migration assay. Gene expression of tenocyte markers, cytokines, growth factors, and matrix metalloprotease (MMP) in tenocytes were assessed using quantitative polymerase chain reaction.
Although A-CTM supported more tenocyte adhesion, I-CTM promoted significantly more tenocyte proliferation compared with A-CTM and B-CTM. Unlike A-CTM, tenocyte migration was higher in I-CTM than the control. The presence of I-CTM also prevented the loss of tenocyte phenotype, attenuated the expression of pro-inflammatory cytokines, growth factors, and MMP, and promoted the expression of antifibrotic growth factor, TGFβ3.
Compared with A-CTM and B-CTM, I-CTM interacted more favorably with human tenocytes in vitro. I-CTM supported tenocyte proliferation with reduced de-differentiation and attenuation of the inflammatory response, suggesting that I-CTM may support tendon healing and regeneration in vivo.
可注射结缔组织基质(CTM)具有微创特性、结构和生化细胞外基质成分以及填充不规则空间的能力,可能促进肌腱愈合。本研究的目的是评估胎盘CTM对人肌腱细胞细胞活性的影响。脱细胞处理,即从CTM中去除细胞、细胞碎片和DNA,已被证明可减少宿主的炎症反应。因此,作者假设脱细胞CTM将提供一个对细胞更友好的基质来支持肌腱细胞功能。
选择三种人胎盘CTM进行比较:AmnioFill®(A-CTM),一种经过最少处理的无活力细胞颗粒;BioRenew™(B-CTM),一种液体基质;以及Interfyl®(I-CTM),一种脱细胞可流动颗粒。使用细胞活力测定评估黏附与增殖,使用Transwell迁移测定评估肌腱细胞迁移。使用定量聚合酶链反应评估肌腱细胞中肌腱细胞标志物、细胞因子、生长因子和基质金属蛋白酶(MMP)的基因表达。
尽管A-CTM支持更多的肌腱细胞黏附,但与A-CTM和B-CTM相比,I-CTM显著促进了更多的肌腱细胞增殖。与A-CTM不同,I-CTM中的肌腱细胞迁移高于对照组。I-CTM的存在还防止了肌腱细胞表型的丧失,减弱了促炎细胞因子、生长因子和MMP的表达,并促进了抗纤维化生长因子TGFβ3的表达。
与A-CTM和B-CTM相比,I-CTM在体外与人类肌腱细胞的相互作用更有利。I-CTM支持肌腱细胞增殖,减少去分化并减轻炎症反应,表明I-CTM可能在体内支持肌腱愈合和再生。