Cartilage engineering using Infrapatellar fat-pad derived MSCs for Osteoarthritis
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摘要Purpose (the aim of the study): Infrapatellar Fat Pad derived MSCs (IFP-MSCs) is an ideal alternative cell source for osteoarthritis related tissue engineering in the fabrication of cartilage grafts or osteochondral constructs. IFP-MSCs could be harvested with ease at large quantities using relatively painless arthroscopic procedures and maintains its chondrogenic capacity in disease conditions such as OA, with no age-dependent decline in cell proliferation and differentiation potential. In this study, IFP-MSCs were differentiated towards chondrogenic lineages using proprietary collagen microencapsulation technology developed from our laboratory and is subsequently fabricated into engineered cartilage tissues for the development of a future OA therapeutic option.
Methods: Human IFP-MSCs (hIFP-MSCs) and Rabbit IFP-MSCs (rIFP-MSCs) were isolated from excised infrapatellar fat pad of OA patients undergoing total knee replacement and New Zealand white rabbits respectively. Experimental approach was classified into two stages, the initial stage begins the optimization of collagen microencapsulation concentration and aggregation number for chondrogenic differentiation, and the second stage involves the fabrication of engineered cartilage using optimized conditions developed from stage 1. In brief, hIFP-MSCs were differentiated for 21 days towards chondrogenic lineage via collagen microencapsulation at a collagen concentration range of 0.05-2mg/ml with an aggregation number between 50-700 to form cartilage microtissues at stage 1, histological, biochemical and biomechanical evaluations of different markers were conducted on the differentiated cartilage microtissue to depict the optimised differentiation conditions of the development of engineered cartilage. Engineered cartilage was subsequently fabricated in stage 2 using cartilage microtissues, the quality of the tissues was assessed using histology and immunohistochemistry.
Results: hIFP-MSCs and rIFP-MSCs microencapsulated with a collagen concentration of 0.05mg/ml and 0.1mg/ml respectively resulted in optimal differentiated cartilage microtissues; Safranin O staining displayed extensive GAG deposition and cartilaginous matrix formation with high expressions of chondrogenic markers including aggrecan and Collagen type-II from immunohistochemistry. Upregulation of chondrogenic gene markers of Sox9 and Collagen type-II were observed and GAG/HYP ratio were measured at a native-like 1.7-2.8:1 for hIFP-MSCs derived cartilage microtissues. Engineered cartilage of both cell types also demonstrated identical findings in histological and immunohistochemical assessments.
Conclusions: IFP-MSCs could be expanded with ease and achieve successful chondrogenic differentiation via collagen microencapsulation to biomimic mesenchymal condensation. Engineered cartilage tissues were successfully fabricated using hIFP-MSC or rIFP-MSCs. IFP-MSC holds tremendous potential in becoming a viable alternative MSC source for OA-related osteochondral and cartilage tissue engineering as future OA therapeutics.
Methods: Human IFP-MSCs (hIFP-MSCs) and Rabbit IFP-MSCs (rIFP-MSCs) were isolated from excised infrapatellar fat pad of OA patients undergoing total knee replacement and New Zealand white rabbits respectively. Experimental approach was classified into two stages, the initial stage begins the optimization of collagen microencapsulation concentration and aggregation number for chondrogenic differentiation, and the second stage involves the fabrication of engineered cartilage using optimized conditions developed from stage 1. In brief, hIFP-MSCs were differentiated for 21 days towards chondrogenic lineage via collagen microencapsulation at a collagen concentration range of 0.05-2mg/ml with an aggregation number between 50-700 to form cartilage microtissues at stage 1, histological, biochemical and biomechanical evaluations of different markers were conducted on the differentiated cartilage microtissue to depict the optimised differentiation conditions of the development of engineered cartilage. Engineered cartilage was subsequently fabricated in stage 2 using cartilage microtissues, the quality of the tissues was assessed using histology and immunohistochemistry.
Results: hIFP-MSCs and rIFP-MSCs microencapsulated with a collagen concentration of 0.05mg/ml and 0.1mg/ml respectively resulted in optimal differentiated cartilage microtissues; Safranin O staining displayed extensive GAG deposition and cartilaginous matrix formation with high expressions of chondrogenic markers including aggrecan and Collagen type-II from immunohistochemistry. Upregulation of chondrogenic gene markers of Sox9 and Collagen type-II were observed and GAG/HYP ratio were measured at a native-like 1.7-2.8:1 for hIFP-MSCs derived cartilage microtissues. Engineered cartilage of both cell types also demonstrated identical findings in histological and immunohistochemical assessments.
Conclusions: IFP-MSCs could be expanded with ease and achieve successful chondrogenic differentiation via collagen microencapsulation to biomimic mesenchymal condensation. Engineered cartilage tissues were successfully fabricated using hIFP-MSC or rIFP-MSCs. IFP-MSC holds tremendous potential in becoming a viable alternative MSC source for OA-related osteochondral and cartilage tissue engineering as future OA therapeutics.
著者Ho Kwan Jeffrey Leung, Chun Hoi Yan, Tim Yun Michael Ong, Shu Hang Patrick Yung, Barbara P. Chan
期刊名稱Osteoarthritis and Cartilage
出版年份2024
月份4
卷號32
期次Suppl 1
出版社Elsevier
頁次S314 - S314
國際標準期刊號1063-4584
電子國際標準期刊號1522-9653
語言美式英語
