BACKGROUND: Radial tears of the meniscus are a common knee injury, frequently resulting in osteoarthritis. To date, there are no established, effective treatments for radial tears. Adipose-derived stem cells (ASCs) may be an attractive cell source for meniscal regeneration because they can be quickly isolated in large number and are capable of undergoing induced fibrochondrogenic differentiation mediated by transforming growth factor β3 (TGF-β3). However, the use of ASCs for meniscal repair is largely unexplored. HYPOTHESIS: ASC-seeded hydrogels with preloaded TGF-β3 will improve meniscal healing of radial tears, as modeled in an explant model. STUDY DESIGN: Controlled laboratory study. METHODS: With an institutional review board-exempted protocol, human ASCs were isolated from the infrapatellar fat pads of 3 donors, obtained after total knee replacement, and characterized. ASCs were encapsulated in photocrosslinkable methacrylated gelatin hydrogels to form 3-dimensional constructs, which were placed into tissue culture. The effect of TGF-β3-whether preloaded into the hydrogel or added as a soluble medium supplement-on matrix-sulfated proteoglycan deposition in the constructs was evaluated. A meniscal explant culture model was used to simulate meniscal repair. Cylindrical-shaped explants were excised from the inner avascular region of adult bovine menisci, and a radial tear was modeled by cutting perpendicular to the meniscal main fibers to the length of the radius. Six combinations of hydrogels-namely, acellular and ASC-seeded hydrogels supplemented with preloaded TGF-β3 (2 µg/mL) or soluble TGF-β3 (10 ng/mL) and without supplement-were injected into the radial tear and stabilized by photocrosslinking with visible light. At 4 and 8 weeks of culture, healing was assessed through histology, immunofluorescence staining, and mechanical testing. RESULTS: ASCs isolated from the 3 donors exhibited colony-forming and multilineage differentiation potential. Hydrogels preloaded with TGF-β3 and those cultured in soluble TGF-β3 showed robust matrix-sulfated proteoglycan deposition. ASC-seeded hydrogels promoted superior healing as compared with acellular hydrogels, with preloaded or soluble TGF-β3 further improving histological scores and mechanical properties. CONCLUSION: These findings demonstrated that ASC-seeded hydrogels preloaded with TGF-β3 enhanced healing of radial meniscal tears in an in vitro meniscal repair model. CLINICAL RELEVANCE: Injection delivery of ASCs in a TGF-β3-preloaded photocrosslinkable hydrogel represents a novel candidate strategy to repair meniscal radial tears and minimize further osteoarthritic joint degeneration.
BACKGROUND: Radial tears of the meniscus are a common knee injury, frequently resulting in osteoarthritis. To date, there are no established, effective treatments for radial tears. Adipose-derived stem cells (ASCs) may be an attractive cell source for meniscal regeneration because they can be quickly isolated in large number and are capable of undergoing induced fibrochondrogenic differentiation mediated by transforming growth factor β3 (TGF-β3). However, the use of ASCs for meniscal repair is largely unexplored. HYPOTHESIS: ASC-seeded hydrogels with preloaded TGF-β3 will improve meniscal healing of radial tears, as modeled in an explant model. STUDY DESIGN: Controlled laboratory study. METHODS: With an institutional review board-exempted protocol, human ASCs were isolated from the infrapatellar fat pads of 3 donors, obtained after total knee replacement, and characterized. ASCs were encapsulated in photocrosslinkable methacrylated gelatin hydrogels to form 3-dimensional constructs, which were placed into tissue culture. The effect of TGF-β3-whether preloaded into the hydrogel or added as a soluble medium supplement-on matrix-sulfated proteoglycan deposition in the constructs was evaluated. A meniscal explant culture model was used to simulate meniscal repair. Cylindrical-shaped explants were excised from the inner avascular region of adult bovine menisci, and a radial tear was modeled by cutting perpendicular to the meniscal main fibers to the length of the radius. Six combinations of hydrogels-namely, acellular and ASC-seeded hydrogels supplemented with preloaded TGF-β3 (2 µg/mL) or soluble TGF-β3 (10 ng/mL) and without supplement-were injected into the radial tear and stabilized by photocrosslinking with visible light. At 4 and 8 weeks of culture, healing was assessed through histology, immunofluorescence staining, and mechanical testing. RESULTS: ASCs isolated from the 3 donors exhibited colony-forming and multilineage differentiation potential. Hydrogels preloaded with TGF-β3 and those cultured in soluble TGF-β3 showed robust matrix-sulfated proteoglycan deposition. ASC-seeded hydrogels promoted superior healing as compared with acellular hydrogels, with preloaded or soluble TGF-β3 further improving histological scores and mechanical properties. CONCLUSION: These findings demonstrated that ASC-seeded hydrogels preloaded with TGF-β3 enhanced healing of radial meniscal tears in an in vitro meniscal repair model. CLINICAL RELEVANCE: Injection delivery of ASCs in a TGF-β3-preloaded photocrosslinkable hydrogel represents a novel candidate strategy to repair meniscal radial tears and minimize further osteoarthritic joint degeneration.
Authors: Heenam Kwon; Wendy E Brown; Cassandra A Lee; Dean Wang; Nikolaos Paschos; Jerry C Hu; Kyriacos A Athanasiou Journal: Nat Rev Rheumatol Date: 2019-07-11 Impact factor: 20.543
Authors: Sofia Hidalgo Perea; Lucas P Lyons; James F Nishimuta; J Brice Weinberg; Amy L McNulty Journal: Connect Tissue Res Date: 2019-10-29 Impact factor: 3.417
Authors: Meagan J Makarczyk; Qi Gao; Yuchen He; Zhong Li; Michael S Gold; Mark C Hochberg; Bruce A Bunnell; Rocky S Tuan; Stuart B Goodman; Hang Lin Journal: Tissue Eng Part C Methods Date: 2021-02-04 Impact factor: 3.056