Ye Ken, Traianedes Kathy, Choong Peter F M, Myers Damian E
Department of Surgery, St Vincent's Hospital Melbourne, The University of Melbourne, Fitzroy, VIC, Australia; Department of Orthopaedics, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.
Department of Clinical Neurosciences, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia; Department of Medicine, St Vincent's Hospital Melbourne, The University of Melbourne, Fitzroy, VIC, Australia.
Front Surg. 2016 Jan 26;3:3. doi: 10.3389/fsurg.2016.00003. eCollection 2016.
Acellular dermal matrix (ADM) has been in clinical use for decades in numerous surgical applications. The ability for ADM to promote cellular repopulation, revascularisation and tissue regeneration is well documented. Adipose stem cells have the ability to differentiate into mesenchymal tissue types, including bone and cartilage. The aim of this study was to investigate the potential interaction between ADM and adipose stem cells in vitro using TGFβ3 and BMP6. Human infrapatellar fat pad-derived adipose stem cells (IPFP-ASC) were cultured with ADM derived from rat dermis in chondrogenic (TGFβ3 and BMP6) medium in vitro for 2 and 4 weeks. Histology, qPCR, and immunohistochemistry were performed to assess for markers of chondrogenesis (collagen Type II, SOX9 and proteoglycans). At 4 weeks, cell-scaffold constructs displayed cellular changes consistent with chondrogenesis, with evidence of stratification of cell layers and development of a hyaline-like cartilage layer superficially, which stained positively for collagen Type II and proteoglycans. Significant cell-matrix interaction was seen between the cartilage layer and the ADM itself with seamless integration between each layer. Real time qPCR showed significantly increased COL2A1, SOX9, and ACAN gene expression over 4 weeks when compared to control. COL1A2 gene expression remained unchanged over 4 weeks. We believe that the principles that make ADM versatile and successful for tissue regeneration are applicable to cartilage regeneration. This study demonstrates in vitro the ability for IPFP-ASCs to undergo chondrogenesis, infiltrate, and interact with ADM. These outcomes serve as a platform for in vivo modelling of ADM for cartilage repair.
脱细胞真皮基质（ADM）已在众多外科手术应用中临床使用了数十年。ADM促进细胞再填充、血管再生和组织再生的能力已有充分记录。脂肪干细胞具有分化为间充质组织类型的能力，包括骨和软骨。本研究的目的是使用转化生长因子β3（TGFβ3）和骨形态发生蛋白6（BMP6）在体外研究ADM与脂肪干细胞之间的潜在相互作用。人髌下脂肪垫来源的脂肪干细胞（IPFP-ASC）在体外与大鼠真皮来源的ADM一起在软骨形成（TGFβ3和BMP6）培养基中培养2周和4周。进行组织学、定量聚合酶链反应（qPCR）和免疫组织化学以评估软骨形成标志物（II型胶原蛋白、SOX9和蛋白聚糖）。在4周时，细胞-支架构建体显示出与软骨形成一致的细胞变化，有细胞层分层的证据，并且表面有类似透明软骨层的形成，该层对II型胶原蛋白和蛋白聚糖染色呈阳性。在软骨层和ADM本身之间观察到显著的细胞-基质相互作用，各层之间无缝整合。实时qPCR显示，与对照相比，4周内COL2A1、SOX9和ACAN基因表达显著增加。COL1A2基因表达在4周内保持不变。我们认为，使ADM在组织再生中具有通用性和成功性的原理适用于软骨再生。本研究在体外证明了IPFP-ASC进行软骨形成、浸润并与ADM相互作用的能力。这些结果为ADM用于软骨修复的体内建模提供了一个平台。
