Hitomi Sano1, Hakan Orbay2, Hiroto Terashi3, Hiko Hyakusoku2, Rei Ogawa2. 1. Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan; Department of Surgical Science, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. Electronic address: sasasa116sasasa@hotmail.com. 2. Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan. 3. Department of Plastic and Reconstructive Surgery, Kobe University, Kobe, Japan.
Abstract
BACKGROUND: In conventional tissue-regeneration technologies, stem cells and/or other cells are injected into or incubated on scaffolds. In general, scaffolds can be classified into synthetic and natural polymers and natural matrices. Polymers are generally less suitable than natural matrices in terms of biocompatibility and biodegradability. A highly promising alternative may be the acellular adipose matrix (AAM), which is a natural scaffold that could mediate tissue regeneration without any artefacts. The optimal method for adipose-tissue decellularisation is described in this article. METHODS: Discarded human adipose tissues harvested from routine operations were used. In experiment 1, four different adipose-tissue-decellularisation methods were compared and modified. In experiment 2, the most effective method was tested by using adipose-tissue blocks from various donor sites (the abdomen, chest and forearm) and of different weights (0.8, 25 and 80 g). Haematoxylin and eosin (H &E) staining, immunohistochemistry (IHC) and scanning electron microscopy were used to determine the efficacy of decellularisation. RESULTS: In experiment 1, a method using an enzymatic digestion solution yielded complete decellularisation after some modifications. In experiment 2, the 0.8-g specimens were completely decellularised by the modified method. However, cell components remained in the 25- and 80-g specimens. The donor site had no effect on the degree of decellularisation. CONCLUSIONS: An optimal method for adipose-tissue decellularisation is reported. Because AAM is a natural collagen scaffold that is of human origin, this report describes an important first step in a tissue-engineering innovation that may be suitable for the regeneration of various tissues.
BACKGROUND: In conventional tissue-regeneration technologies, stem cells and/or other cells are injected into or incubated on scaffolds. In general, scaffolds can be classified into synthetic and natural polymers and natural matrices. Polymers are generally less suitable than natural matrices in terms of biocompatibility and biodegradability. A highly promising alternative may be the acellular adipose matrix (AAM), which is a natural scaffold that could mediate tissue regeneration without any artefacts. The optimal method for adipose-tissue decellularisation is described in this article. METHODS: Discarded human adipose tissues harvested from routine operations were used. In experiment 1, four different adipose-tissue-decellularisation methods were compared and modified. In experiment 2, the most effective method was tested by using adipose-tissue blocks from various donor sites (the abdomen, chest and forearm) and of different weights (0.8, 25 and 80 g). Haematoxylin and eosin (H &E) staining, immunohistochemistry (IHC) and scanning electron microscopy were used to determine the efficacy of decellularisation. RESULTS: In experiment 1, a method using an enzymatic digestion solution yielded complete decellularisation after some modifications. In experiment 2, the 0.8-g specimens were completely decellularised by the modified method. However, cell components remained in the 25- and 80-g specimens. The donor site had no effect on the degree of decellularisation. CONCLUSIONS: An optimal method for adipose-tissue decellularisation is reported. Because AAM is a natural collagen scaffold that is of human origin, this report describes an important first step in a tissue-engineering innovation that may be suitable for the regeneration of various tissues.
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