Ethanol treatment of nanoPGA/PCL composite scaffolds enhances human chondrocyte development in the cellular microenvironment of tissue-engineered auricle constructs
| Autor: | Hirohisa Kusuhara, Noritaka Isogai, Takeshi Teramura, William J. Landis, Shinichi Asamura, Ananth S. Murthy, Yu Sueyoshi, Narihiko Hirano, Robin Jacquet |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Male Scaffold Polymers Cell Culture Techniques Nanofibers Gene Expression Apoptosis 02 engineering and technology Scaffold Seeding Mice Tissue engineering Animal Cells Medicine and Health Sciences Nanotechnology Ear External Materials Immune Response Cells Cultured Connective Tissue Cells Multidisciplinary Cell Death Tissue Scaffolds Chemistry Organic Compounds Cell Differentiation medicine.anatomical_structure Macromolecules Cellular Microenvironment Connective Tissue Cell Processes Physical Sciences Medicine Engineering and Technology Female Anatomy Cellular Types Ear Cartilage Chondrogenesis Research Article Ear Auricle Science 0206 medical engineering Materials Science Immunology Connective tissue Mice Nude Chondrocyte 03 medical and health sciences Chondrocytes Signs and Symptoms Dogs medicine Genetics Animals Humans Cell adhesion Nanomaterials Congenital Microtia Inflammation Ethanol Tissue Engineering Regeneration (biology) Cartilage Organic Chemistry Chemical Compounds Biology and Life Sciences Cell Biology Polymer Chemistry 020601 biomedical engineering 030104 developmental biology Biological Tissue Alcohols Clinical Medicine Polyglycolic Acid Biomedical engineering |
| Zdroj: | PLoS ONE PLoS ONE, Vol 16, Iss 7, p e0253149 (2021) |
| ISSN: | 1932-6203 |
| Popis: | A major obstacle for tissue engineering ear-shaped cartilage is poorly developed tissue comprising cell-scaffold constructs. To address this issue, bioresorbable scaffolds of poly-ε-caprolactone (PCL) and polyglycolic acid nanofibers (nanoPGA) were evaluated using an ethanol treatment step before auricular chondrocyte scaffold seeding, an approach considered to enhance scaffold hydrophilicity and cartilage regeneration. Auricular chondrocytes were isolated from canine ears and human surgical samples discarded during otoplasty, including microtia reconstruction. Canine chondrocytes were seeded onto PCL and nanoPGA sheets either with or without ethanol treatment to examine cellular adhesion in vitro. Human chondrocytes were seeded onto three-dimensional bioresorbable composite scaffolds (PCL with surface coverage of nanoPGA) either with or without ethanol treatment and then implanted into athymic mice for 10 and 20 weeks. On construct retrieval, scanning electron microscopy showed canine auricular chondrocytes seeded onto ethanol-treated scaffolds in vitro developed extended cell processes contacting scaffold surfaces, a result suggesting cell-scaffold adhesion and a favorable microenvironment compared to the same cells with limited processes over untreated scaffolds. Adhesion of canine chondrocytes was statistically significantly greater (p ≤ 0.05) for ethanol-treated compared to untreated scaffold sheets. After implantation for 10 weeks, constructs of human auricular chondrocytes seeded onto ethanol-treated scaffolds were covered with glossy cartilage while constructs consisting of the same cells seeded onto untreated scaffolds revealed sparse connective tissue and cartilage regeneration. Following 10 weeks of implantation, RT-qPCR analyses of chondrocytes grown on ethanol-treated scaffolds showed greater expression levels for several cartilage-related genes compared to cells developed on untreated scaffolds with statistically significantly increased SRY-box transcription factor 5 (SOX5) and decreased interleukin-1α (inflammation-related) expression levels (p ≤ 0.05). Ethanol treatment of scaffolds led to increased cartilage production for 20- compared to 10-week constructs. While hydrophilicity of scaffolds was not assessed directly in the present findings, a possible factor supporting the summary data is that hydrophilicity may be enhanced for ethanol-treated nanoPGA/PCL scaffolds, an effect leading to improvement of chondrocyte adhesion, the cellular microenvironment and cartilage regeneration in tissue-engineered auricle constructs. |
| Databáze: | OpenAIRE |
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