Sanders Mitchell Corey, Balaji Swathi, Martin W Blake, Siegmund Nikole, Poland Lindsay, Sanders Hanna Mia, Wei Da, Kaliada Hanna, Littlejohn Sara, Ganey Timothy
ProDevLab, Alira Health, Framingham, Massachusetts, USA.
Pediatric Surgery Division, Department of Surgery, Baylor College of Medicine and Texas Children's Hospital, Houston, Texas, USA.
Wound Repair Regen. 2023 Jul-Aug;31(4):475-488. doi: 10.1111/wrr.13099. Epub 2023 Jun 26.
Recent evidence suggests that protecting human amnion and chorion matrices (HACM) during processing enhances the performance of HACM for wound repair and tissue regeneration. We utilised a diabetic (db/db) delayed wound healing mouse model. Treatment of db/db full-thickness excisional wounds with HACM, processed with a polyampholyte preservative accentuated the proliferative phase of wound healing that decreased the time necessary to heal wounds. Polyampholyte protection improved the preservation of growth factors and cytokines during room temperature storage following E-beam sterilisation and improved its function in wound healing applications. Our findings indicate protected HACM tissue up-regulated MIP2, NF-kB, TNF-α, KI-67, and Arg1 (0.6-fold to 1.5-fold) but those changes were not statistically significant. Immunofluorescent assessment identifying cell activity illustrated an induction of the proliferative phase of wound healing and a switch from an inflammatory macrophage phenotype (M1) to a pro-regenerative macrophage phenotype (M2a). Genomic profiling of 282 genes was performed using Nanostring from co-cultures of human macrophages and fibroblasts. The polyampholyte + HACM-treated group, compared with the HACM or polyampholyte alone groups, had a statistically significant up-regulation (32-368 fold) of 12 genes primarily involved in macrophage plasticity including CLC7, CD209, CD36, HSD11B1, ICAM1, IL1RN, IL3RA, ITGAX, LSP1, and PLXDC2 (adj. p-value < 0.05). The polyampholyte alone group demonstrated statistically significant down-regulation of four genes ADRA2, COL7A1, CSF3, and PTGS2 (adj. p < 0.05). The HACM alone group up-regulated four genes ATG14, CXCL11, DNMT3A, and THBD, but the results were not statistically significant. Biomechanical measurements indicated that wounds treated with polyampholyte-protected HACM had more tensile integrity compared with wounds treated with HACM alone. These findings indicate that better protection of HACM during processing stabilises the HACM matrix, which may lead to improved wound healing outcomes.
近期证据表明，在处理过程中保护人羊膜和绒毛膜基质（HACM）可提高其在伤口修复和组织再生方面的性能。我们使用了糖尿病（db/db）延迟伤口愈合小鼠模型。用聚两性电解质防腐剂处理的HACM治疗db/db全层切除伤口，可加速伤口愈合的增殖期，减少伤口愈合所需时间。聚两性电解质保护改善了电子束灭菌后室温储存期间生长因子和细胞因子的保存，并改善了其在伤口愈合应用中的功能。我们的研究结果表明，受保护的HACM组织上调了MIP2、NF-kB、TNF-α、KI-67和Arg1（0.6倍至1.5倍），但这些变化无统计学意义。免疫荧光评估确定细胞活性显示伤口愈合增殖期的诱导以及从炎性巨噬细胞表型（M1）向促再生巨噬细胞表型（M2a）的转变。使用Nanostring对人巨噬细胞和成纤维细胞共培养物进行了282个基因的基因组分析。与单独使用HACM或聚两性电解质组相比，聚两性电解质+HACM处理组有12个主要参与巨噬细胞可塑性的基因出现统计学显著上调（32-368倍），包括CLC7、CD209、CD36、HSD11B1、ICAM1、IL1RN、IL3RA、ITGAX、LSP1和PLXDC2（校正p值<0.05）。单独使用聚两性电解质组有四个基因ADRA2、COL7A1、CSF3和PTGS2出现统计学显著下调（校正p<0.05）。单独使用HACM组上调了四个基因ATG14、CXCL11、DNMT3A和THBD，但结果无统计学意义。生物力学测量表明，与单独使用HACM处理的伤口相比，用聚两性电解质保护的HACM处理的伤口具有更高的拉伸完整性。这些发现表明，在处理过程中更好地保护HACM可稳定HACM基质，这可能会改善伤口愈合结果。
