Second-harmonic generation microscopy for assessment of mesenchymal stem cell-seeded acellular dermal matrix in wound-healing |
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| Affiliation: | 1. MOE Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, Guangdong, China;2. Research Resources Center, South China Normal University, Guangzhou, Guangdong, China;3. Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong, China;1. Department of Genetics, University of Rzeszow, Rejtana 16C, 35-959 Rzeszow, Poland;2. Department of Plant Physiology, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa, Poland;3. Department of Biochemistry and Cell Biology, University of Rzeszow, Zelwerowicza 4, 35-601 Rzeszow, Poland;1. Institute of Tissue Regeneration Engineering (ITREN) and Department of Biomaterials Science, School of Dentistry, Dankook University, Cheonan 330-714, Republic of Korea;2. Department of Nanobiomedical Science & BK21 PLUS NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 330-714, Republic of Korea;3. Department of Advanced Materials, Hannam University, Daejeon 305-811, Republic of Korea;1. Medical School & State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China;2. Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing 210008, China;3. Institute of Urology, Nanjing University, Nanjing 210008, China;1. Department of Anesthesiology, Yale University, New Haven, CT 06519, USA;2. Department Biomedical Engineering, Yale University, New Haven, CT 06519, USA;3. Department of Medicine, Section of Cardiovascular Medicine, Yale University, New Haven, CT 06519, USA;1. Division of Dermatology and Venereology, Department of Clinical Sciences, Lund University, Biomedical Center, Tornavägen 10, SE-22184 Lund, Sweden;2. Division of Respiratory Medicine and Allergology, Department of Clinical Sciences, Lund University, Biomedical Center, Tornavägen 10, SE-22184 Lund, Sweden;3. Division of Infection Medicine, Department of Clinical Sciences, Lund University, Biomedical Center, Tornavägen 10, SE-22184 Lund, Sweden;4. Deutsches Institut für Zell- und Gewebeersatz gGmbH, Berlin, Germany;5. Lee Kong Chian School of Medicine, Nanyang Technological University, 308232, Singapore |
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| Abstract: | Direct intra-skin injection of mesenchymal stem cells (MSCs) and the use of biomaterial scaffolds for grafts are both promising approaches of skin wound repair, however they still cannot generate skin that completely resembles the natural skin structures. In this study, we combined these two approaches by using acellular dermal matrix (ADM) recellularized with MSCs to repair cutaneous wounds in a murine model and two-photon fluorescence (TPF) microscopy and second-harmonic generation (SHG) microscopy to assess the effects of this therapy on wound healing. Bone marrow-derived mesenchymal stem cells (BM-MSCs) were tagged with GFP and seeded into ADM (ADM-MSC) via MSC and ADM co-culture. ADM-MSC, ADM or saline was applied to murine excisional skin wounds and wound-healing was evaluated by histological examination on days 7, 14, 21 and TFP microscopy on days 1, 3, 5 and 21 post-treatment. ADM-MSC promoted healing significantly more than treatment with ADM or saline alone, as it led to substantial neovascularization and complete skin appendage regeneration. Furthermore, the SHG microscopic imaging technique proved to be a useful tool for monitoring changes in the collagen network at the wound site during the healing process and assessing the effects of different therapies. |
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| Keywords: | Wound healing Acellular dermal matrix (ADM) Mesenchymal stem cells (MSCs) Two-photon fluorescence (TPF) microscopy Second-harmonic generation (SHG) microscopy |
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