Department of Plastic Surgery and Hand Surgery, University Hospital Zurich, 8091 Zurich, Switzerland.
Center for Preclinical Development, University Hospital Zurich, 8091 Zurich, Switzerland.
Int J Mol Sci. 2023 Sep 11;24(18):13944. doi: 10.3390/ijms241813944.
Mechanically processed stromal vascular fraction (mSVF) is a highly interesting cell source for regenerative purposes, including wound healing, and a practical alternative to enzymatically isolated SVF. In the clinical context, SVF benefits from scaffolds that facilitate viability and other cellular properties. In the present work, the feasibility of methacrylated gelatin (GelMA), a stiffness-tunable, light-inducible hydrogel with high biocompatibility is investigated as a scaffold for SVF in an in vitro setting. Lipoaspirates from elective surgical procedures were collected and processed to mSVF and mixed with GelMA precursor solutions. Non-encapsulated mSVF served as a control. Viability was measured over 21 days. Secreted basic fibroblast growth factor (bFGF) levels were measured on days 1, 7 and 21 by ELISA. IHC was performed to detect VEGF-A, perilipin-2, and CD73 expression on days 7 and 21. The impact of GelMA-mSVF on human dermal fibroblasts was measured in a co-culture assay by the same viability assay. The viability of cultured GelMA-mSVF was significantly higher after 21 days ( < 0.01) when compared to mSVF alone. Also, GelMA-mSVF secreted stable levels of bFGF over 21 days. While VEGF-A was primarily expressed on day 21, perilipin-2 and CD73-positive cells were observed on days 7 and 21. Finally, GelMA-mSVF significantly improved fibroblast viability as compared with GelMA alone ( < 0.01). GelMA may be a promising scaffold for mSVF as it maintains cell viability and proliferation with the release of growth factors while facilitating adipogenic differentiation, stromal cell marker expression and fibroblast proliferation.
机械加工的基质血管成分(mSVF)是一种非常有前途的用于再生目的的细胞来源,包括伤口愈合,并且是酶分离的 SVF 的实用替代品。在临床环境中,SVF 受益于促进细胞活力和其他细胞特性的支架。在本工作中,研究了具有高生物相容性的光诱导的可调节刚度甲基丙烯酰化明胶(GelMA)作为体外 SVF 支架的可行性。从选择性手术中收集和处理脂肪抽吸物以获得 mSVF,并将其与 GelMA 前体溶液混合。非包封的 mSVF 作为对照。在 21 天内测量活力。通过 ELISA 在第 1、7 和 21 天测量分泌的碱性成纤维细胞生长因子(bFGF)水平。在第 7 和 21 天通过免疫组织化学(IHC)检测 VEGF-A、 perilipin-2 和 CD73 的表达。通过相同的活力测定,在共培养测定中测量 GelMA-mSVF 对人真皮成纤维细胞的影响。与单独的 mSVF 相比,培养的 GelMA-mSVF 在 21 天后的活力显着更高(<0.01)。此外,GelMA-mSVF 在 21 天内稳定分泌 bFGF。虽然 VEGF-A 主要在第 21 天表达,但在第 7 和 21 天观察到 perilipin-2 和 CD73 阳性细胞。最后,与单独的 GelMA 相比,GelMA-mSVF 显着提高了成纤维细胞的活力(<0.01)。GelMA 可能是 mSVF 的一种有前途的支架,因为它在释放生长因子的同时维持细胞活力和增殖,同时促进脂肪生成分化、基质细胞标志物表达和成纤维细胞增殖。