DataSheet1_Assessment of inflammatory suppression and fibroblast infiltration in tissue remodelling by supercritical CO2 acellular dermal matrix (scADM) utilizing Sprague Dawley models.ZIP

Human skin-derived ECM aids cell functions but can trigger immune reactions; therefore it is addressed through decellularization. Acellular dermal matrices (ADMs), known for their regenerative properties, are used in tissue and organ regeneration. ADMs now play a key role in plastic and reconstructive surgery, enhancing aesthetics and reducing capsular contracture risk. Innovative decellularization with supercritical carbon dioxide preserves ECM quality for clinical use. The study investigated the cytotoxicity, biocompatibility, and anti-inflammatory properties of supercritical CO2 acellular dermal matrix (scADM) in vivo based on Sprague Dawley rat models. Initial experiments in vitro with fibroblast cells confirmed the non-toxic nature of scADM and demonstrated cell infiltration into scADMs after incubation. Subsequent tests in vitro revealed the ability of scADM to suppress inflammation induced by lipopolysaccharides (LPS) presenting by the reduction of pro-inflammatory cytokines TNF-α, IL-6, IL-1β, and MCP-1. In the in vivo model, histological assessment of implanted scADMs in 6 months revealed a decrease in inflammatory cells, confirmed further by the biomarkers of inflammation in immunofluorescence staining. Besides, an increase in fibroblast infiltration and collagen formation was observed in histological staining, which was supported by various biomarkers of fibroblasts. Moreover, the study demonstrated vascularization and macrophage polarization, depicting increased endothelial cell formation. Alteration of matrix metalloproteinases (MMPs) was analyzed by RT-PCR, indicating the reduction of MMP2, MMP3, and MMP9 levels over time. Simultaneously, an increase in collagen deposition of collagen I and collagen III was observed, verified in immunofluorescent staining, RT-PCR, and western blotting. Overall, the findings suggested that scADMs offer significant benefits in improving outcomes in implant-based procedures as well as soft tissue substitution.

源自人类皮肤的细胞外基质（Extracellular Matrix，ECM）可辅助细胞发挥功能，但亦可能引发免疫反应，因此需通过脱细胞技术对其进行处理。脱细胞真皮基质（Acellular Dermal Matrices，ADMs）因具备再生特性，被应用于组织与器官再生领域。如今，ADMs已在整形与重建外科中发挥关键作用，可提升美学修复效果并降低包膜挛缩风险。采用超临界二氧化碳的创新脱细胞技术能够保留ECM的临床应用品质。本研究以斯普拉-道来大鼠（Sprague Dawley rat）为实验模型，在体内水平探究了超临界二氧化碳脱细胞真皮基质（supercritical CO₂ acellular dermal matrix，scADM）的细胞毒性、生物相容性与抗炎特性。体外成纤维细胞实验初步验证了scADM无细胞毒性，并证实孵育后有成纤维细胞向scADM内部浸润。后续体外实验表明，scADM可通过下调促炎细胞因子TNF-α、IL-6、IL-1β及MCP-1的表达，抑制脂多糖（Lipopolysaccharides，LPS）诱导的炎症反应。在体内模型中，对植入6个月的scADM进行组织学评估，发现炎症细胞浸润减少，这一结果通过免疫荧光染色的炎症生物标志物得到进一步证实。此外，组织学染色观察到成纤维细胞浸润与胶原形成均有所增加，该现象得到多种成纤维细胞生物标志物的佐证。本研究还证实了血管生成与巨噬细胞极化现象，表现为内皮细胞形成增多。本研究通过逆转录聚合酶链反应（RT-PCR）分析了基质金属蛋白酶（Matrix Metalloproteinases，MMPs）的表达变化，结果显示MMP2、MMP3及MMP9的表达水平随时间推移呈下降趋势。与此同时，Ⅰ型胶原与Ⅲ型胶原的沉积量有所增加，这一结果通过免疫荧光染色、RT-PCR及蛋白质印迹法得到验证。综上，本研究结果表明，scADM在提升植入手术效果及软组织替代治疗方面具有显著优势。