Intraoperative use of enriched collagen and elastin matrices with freshly isolated adipose-derived stem/stromal cells: a potential clinical approach for soft tissue reconstruction

BMC Surgery

Norbert Pallua²  Hans-Oliver Rennekampff²  Michael Vogt³  Christian Opländer²  Sultan Almakadi²  Ziyad Alharbi¹ 

[1] Division of Plastic Surgery, Specialist Surgery Center, King Abdullah Medical City, Mecca, Kingdom of Saudi Arabia;Department of Plastic, Reconstructive and Hand Surgery - Burn Center, Medical Faculty, RWTH Aachen University, Pauwelsstr. 30, Aachen D-52074, Germany;Two-Photon Microscopy Facility, Interdisciplinary Center for Clinical Research (IZKF), Medical Faculty, RWTH Aachen University, Aachen, Germany

关键词: Regeneration and tissue engineering;    Collagen-based scaffolds;    Biomaterials;    Fat grafting;    Liposuction;    Stromal vascular fraction;    Adipose tissue-derived stem/stromal cells;   

Others  :  866882
DOI  :  10.1186/1471-2482-14-10

 received in 2013-05-03, accepted in 2014-02-17,  发布年份 2014

【 摘 要 】

Background

Adipose tissue contains a large number of multipotent cells, which are essential for stem cell-based therapies. The combination of this therapy with suitable commercial clinically used matrices, such as collagen and elastin matrices (i.e. dermal matrices), is a promising approach for soft tissue reconstruction. We previously demonstrated that the liposuction method affects the adherence behaviour of freshly isolated adipose-derived stem/stromal cells (ASCs) on collagen and elastin matrices. However, it remains unclear whether freshly isolated and uncultured ASCs could be directly transferred to matrices during a single transplantation operation without additional cell culture steps.

Methods

After each fat harvesting procedure, ASCs were isolated and directly seeded onto collagen and elastin matrices. Different time intervals (i.e. 1, 3 and 24 h) were investigated to determine the time interval needed for cellular attachment to the collagen and elastin matrices. Resazurin-based vitality assays were performed after seeding the cells onto the collagen and elastin matrices. In addition, the adhesion and migration of ASCs on the collagen and elastin matrices were visualised using histology and two-photon microscopy.

Results

A time-dependent increase in the number of viable ASCs attached to the collagen and elastin matrices was observed. This finding was supported by mitochondrial activity and histology results. Importantly, the ASCs attached and adhered to the collagen and elastin matrices after only 1 h of ex vivo enrichment. This finding was also supported by two-photon microscopy, which revealed the presence and attachment of viable cells on the upper layer of the construct.

Conclusion

Freshly isolated uncultured ASCs can be safely seeded onto collagen and elastin matrices for ex vivo cellular enrichment of these constructs after liposuction. Although we observed a significant number of seeded cells on the matrices after a 3-h enrichment time, we also observed an adequate number of isolated cells after a 1-h enrichment time. However, this approach must be optimised for clinical use. Thus, in vivo studies and clinical trials are needed to investigate the feasibility of this approach.

【 授权许可】

   
2014 Alharbi et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Fraser JK, Zhu M, Wulur I, Alfonso Z: Adipose-derived stem cells. Methods Mol Biol 2008, 449:59-67.
• [2]Tholpady SS, Llull R, Ogle RC, Rubin JP, Futrell JW, Katz AJ: Adipose tissue: stem cells and beyond. Clin Plast Surg 2006, 33(1):55-62.
• [3]Mizuno H: Adipose-derived stem cells for tissue repair and regeneration: ten years of research and a literature review. J Nihon Med Sch 2009, 76(2):56-66.
• [4]Gimble JM: Adipose tissue-derived therapeutics. Expert Opin Biol Ther 2003, 3:705-713.
• [5]Alharbi Z, Opländer C, Almakadi S, Fritz A, Vogt M, Pallua N: Conventional vs. micro-fat harvesting: how fat harvesting technique affects tissue-engineering approaches using adipose tissue-derived stem/stromal cells. J Plast Reconstr Aesthet Surg 2013, 66(9):1271-1278.
• [6]Keck M, Haluza D, Lumenta DB, Burjak S, Eisenbock B, Kamolz LP, Frey M: Construction of a multi-layer skin substitute: Simultaneous cultivation of keratinocytes and preadipocytes on a dermal template. Burns 2011, 37(4):626-630.
• [7]Coleman SR: Structural fat grafts: the ideal filler? Clin Plast Surg 2001, 28:111-119.
• [8]Chang Q, Li J, Dong Z, Liu L, Lu F: Quantitative volumetric analysis of progressive hemifacial atrophy corrected using stromal vascular fraction-supplemented autologous fat grafts. Dermatol Surg 2013, 39(10):1465-1473.
• [9]Jeon H, Kim J, Yeo H, Jeong H, Son D, Han K: Treatment of diabetic foot ulcer using matriderm in comparison with a skin graft. Arch Plast Surg 2013, 40(4):403-408.
• [10]Walum E, Hansson E, Harvey AL: In vitro testing of neurotoxicity. ATLA 1990, 18:153-179.
• [11]Li K, Wong D, Hiscott P, Stanga P, Groenewald C, McGalliard J: Trypan blue staining of internal limiting membrane and epiretinal membrane during vitrectomy: Visual results and histopathological findings. Br J Ophthalmol 2003, 87:216-219.
• [12]Ahmed SA, Gogal RM Jr, Walsh JE: A new Rapid and Simple Non-Radioactive assay to Monitor and Determine the proliferation of Lymphocytes: An Alternative to H3-thymidine incorporation assay. J lmmunol Methods 1994, 170:211-224.
• [13]Alley MC, Scudiero DA, Monks A, Hursey ML, Czerwinski MJ, Fine DL, Abbott BJ, Mayo JG, Shoemaker RH, Boyd MR: Feasibility of Drug Screening with Panels of Human Tumor Cell Lines Using a Microculture Tetrazolium Assay. Cancer Res 1988, 48:589-601.