Int J Med Sci 2021; 18(10):2217-2227. doi:10.7150/ijms.56342

Research Paper

A novel 3D histotypic cartilage construct engineered by supercritical carbon dioxide decellularized porcine nasal cartilage graft and chondrocytes exhibited chondrogenic capability in vitro

Su-Shin Lee1,2,3,4, Yi-Chia Wu1,2,3, Shu-Hung Huang1,2,3, Ying-Che Chen4, Periasamy Srinivasan5, Dar-Jen Hsieh5, Yi-Chun Yeh5, Yi-Ping Lai5, Yun-Nan Lin1✉

1. Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung City, Taiwan.
2. Department of Surgery, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung City, Taiwan.
3. Regenerative medicine and cell therapy research centre, Kaohsiung Medical University, Kaohsiung, Taiwan.
4. Department of Surgery, Kaohsiung Municipal Siaogang Hospital, Kaohsiung, Taiwan.
5. Center of Research and Development, ACRO Biomedical Co., Ltd. Kaohsiung, Taiwan.

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Citation:
Lee SS, Wu YC, Huang SH, Chen YC, Srinivasan P, Hsieh DJ, Yeh YC, Lai YP, Lin YN. A novel 3D histotypic cartilage construct engineered by supercritical carbon dioxide decellularized porcine nasal cartilage graft and chondrocytes exhibited chondrogenic capability in vitro. Int J Med Sci 2021; 18(10):2217-2227. doi:10.7150/ijms.56342. Available from https://www.medsci.org/v18p2217.htm

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Abstract

Augmentative and reconstructive rhinoplasty surgical procedures use autologous tissue grafts or synthetic grafts to repair the nasal defect and aesthetic reconstruction. Donor site trauma and morbidity are common in autologous grafts. The desperate need for the production of grafted 3D cartilage tissues as rhinoplasty grafts without the adverse effect is the need of the hour. In the present study, we developed a bioactive 3D histotypic construct engineered with the various ratio of adipose-derived stem cells (ADSC) and chondrocytes together with decellularized porcine nasal cartilage graft (dPNCG). We decellularized porcine nasal cartilage using supercritical carbon dioxide (SCCO2) extraction technology. dPNCG was characterized by H&E, DAPI, alcian blue staining, scanning electron microscopy and residual DNA content, which demonstrated complete decellularization. 3D histotypic constructs were engineered using dPNCG, rat ADSC and chondrocytes with different percentage of cells and cultured for 21 days. dPNCG together with 100% chondrocytes produced a solid mass of 3D histotypic cartilage with significant production of glycosaminoglycans. H&E and alcian blue staining showed an intact mass, with cartilage granules bound to one another by extracellular matrix and proteoglycan, to form a 3D structure. Besides, the expression of chondrogenic markers, type II collagen, aggrecan and SOX-9 were elevated indicating chondrocytes cultured on dPNCG substrate facilitates the synthesis of type II collagen along with extracellular matrix to produce 3D histotypic cartilage. To conclude, dPNCG is an excellent substrate scaffold that might offer a suitable environment for chondrocytes to produce 3D histotypic cartilage. This engineered 3D construct might serve as a promising future candidate for cartilage tissue engineering in rhinoplasty.

Keywords: supercritical carbon dioxide, 3D histotypic cartilage, decellularized porcine nasal cartilage graft (dPNCG), chondrocytes, adipose-derived stem cells