Hirsch Tobias, Laemmle Christine, Behr Bjoern, Lehnhardt Marcus, Jacobsen Frank, Hoefer Dirk, Kueckelhaus Maximilian
Department of Plastic Surgery, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum, Germany.
Department of Hygiene, Environment and Medicine, Hohenstein Institutes, Boennigheim, Germany.
J Plast Reconstr Aesthet Surg. 2018 Jan;71(1):101-111. doi: 10.1016/j.bjps.2017.08.009. Epub 2017 Aug 16.
Adipose-derived stem cells represent an interesting option for soft tissue replacement as they are easy to procure and can generate their own blood supply through the production of angiogenic factors. We seeded adipose-derived stem cells on a bioresorbable, biocompatible polymer alginate scaffold to generate autologous soft tissue constructs for repair.
We built and optimized an alginate scaffold and tested its biocompatibility using the MTT assay and its hydration capacity. We then isolated, characterized, and differentiated murine, porcine, and human adipose-derived stem cells. We characterized their angiogenic potential in vitro by VEGF ELISA and HUVEC tube formation assay in traditional cell culture substrate and in the actual three-dimensional scaffold. We assessed the angiogenic potential of adipose-derived stem cell-colonized scaffolds in ovo by chorion allantois membrane angiogenesis assay.
Adipose-derived stem cells differentiated into adipocytes within the alginate scaffolds and demonstrated angiogenic activity. VEGF secretion by adipose-derived stem cells decreased significantly over the 21-day course of adipocyte differentiation in traditional cell culture substrate, but not in scaffolds. Adipose-derived stem cells differentiated for 21 days in scaffolds led to the longest HUVEC tube formation. Scaffolds colonized with adipose-derived stem cells resulted in significantly improved vascularization in ovo.
We demonstrate the feasibility of implant production based on adipose-derived stem cell-colonized alginate scaffolds. The implants demonstrate biocompatibility and promote angiogenesis in vitro and in ovo. Therefore, they provide a combination of essential properties for an implant intended for soft tissue replacement.
脂肪来源干细胞是软组织替代的一个有趣选择,因为它们易于获取,并且可以通过产生血管生成因子来形成自身的血液供应。我们将脂肪来源干细胞接种在一种可生物吸收、生物相容的聚合物藻酸盐支架上,以生成用于修复的自体软组织构建体。
我们构建并优化了藻酸盐支架,并使用MTT法测试其生物相容性及其水合能力。然后,我们分离、鉴定并分化了小鼠、猪和人类的脂肪来源干细胞。我们通过VEGF ELISA以及在传统细胞培养底物和实际三维支架中的人脐静脉内皮细胞(HUVEC)管形成试验,在体外表征了它们的血管生成潜力。我们通过绒毛尿囊膜血管生成试验在鸡胚中评估了脂肪来源干细胞定植支架的血管生成潜力。
脂肪来源干细胞在藻酸盐支架内分化为脂肪细胞,并表现出血管生成活性。在传统细胞培养底物中,脂肪来源干细胞在21天的脂肪细胞分化过程中VEGF分泌显著减少,但在支架中则不然。在支架中分化21天的脂肪来源干细胞导致HUVEC管形成最长。用脂肪来源干细胞定植的支架在鸡胚中导致血管化显著改善。
我们证明了基于脂肪来源干细胞定植藻酸盐支架生产植入物的可行性。这些植入物在体外和鸡胚中表现出生物相容性并促进血管生成。因此,它们为用于软组织替代的植入物提供了重要特性的组合。