Davide Edoardo Bonasia1, Antongiulio Marmotti2, Silvia Mattia3, Aurelio Cosentino4, Simone Spolaore4, Giorgio Governale4, Filippo Castoldi4, Roberto Rossi2. 1. Department of Orthopaedics and Traumatology, University of Torino, CTO Hospital, Torino, Italy. Electronic address: davidebonasia@virgilio.it. 2. Department of Orthopaedics and Traumatology, University of Torino, Mauriziano "Umberto I" Hospital, Torino, Italy. 3. Molecular Biology Center, University of Torino, Torino, Italy. 4. Department of Orthopaedics and Traumatology, University of Torino, CTO Hospital, Torino, Italy.
Abstract
PURPOSE: To evaluate if the degree of chondral fragmentation affected extracellular matrix (ECM) production in cartilage fragment autograft implantation in vitro. METHODS: Cartilage was taken from 5 donors undergoing total hip replacement (mean age, 65.6 years; standard deviation [SD], 3). The cartilage was minced to obtain 4 groups with different fragment sizes: (1) "fish scale" (diameter, 8 mm; thickness, 0.3 mm), (2) cubes with 2-mm sides, (3) cubes with 1-mm sides, and (4) cartilage paste (< 0.3 mm). The cultures were maintained in chondrogenic medium for 6 weeks. Biochemically, a proteoglycan (PG):DNA ratio was calculated as the best approximation of ECM production per cell. The ratio between PG released in the culture medium and the PG in the neocartilage (PGrel:PG) was used as a matrix stability index. Histologically, the slides were stained with safranin O fast green and collagen type II immunostaining. The titration of safranin O-positive cells and the Bern score were calculated. RESULTS: Regarding the PG:DNA ratio, group 4 performed significantly better than groups 1 (P = .001) and 3 (P = .02), whereas group 2 performed better than group 1 (P = .03). No significant difference was found regarding the PGrel:PG ratio and safranin O-positive cells. Regarding the Bern score, group 4 performed significantly better than groups 1 (P = .02), 2 (P = .04), and 3 (P = .03). CONCLUSIONS: We conclude that human cartilage fragmentation significantly affects ECM production in vitro. Increased fragmentation enhances ECM production. CLINICAL RELEVANCE: Assuming a similar behavior in vivo, we recommend mincing the cartilage into small pieces when performing the cartilage fragment autograft implantation technique in order to increase ECM production. Further in vitro studies investigating cartilage of younger nonarthritic donors, as well as in vivo studies, are needed.
PURPOSE: To evaluate if the degree of chondral fragmentation affected extracellular matrix (ECM) production in cartilage fragment autograft implantation in vitro. METHODS:Cartilage was taken from 5 donors undergoing total hip replacement (mean age, 65.6 years; standard deviation [SD], 3). The cartilage was minced to obtain 4 groups with different fragment sizes: (1) "fish scale" (diameter, 8 mm; thickness, 0.3 mm), (2) cubes with 2-mm sides, (3) cubes with 1-mm sides, and (4) cartilage paste (< 0.3 mm). The cultures were maintained in chondrogenic medium for 6 weeks. Biochemically, a proteoglycan (PG):DNA ratio was calculated as the best approximation of ECM production per cell. The ratio between PG released in the culture medium and the PG in the neocartilage (PGrel:PG) was used as a matrix stability index. Histologically, the slides were stained with safranin O fast green and collagen type II immunostaining. The titration of safranin O-positive cells and the Bern score were calculated. RESULTS: Regarding the PG:DNA ratio, group 4 performed significantly better than groups 1 (P = .001) and 3 (P = .02), whereas group 2 performed better than group 1 (P = .03). No significant difference was found regarding the PGrel:PG ratio and safranin O-positive cells. Regarding the Bern score, group 4 performed significantly better than groups 1 (P = .02), 2 (P = .04), and 3 (P = .03). CONCLUSIONS: We conclude that humancartilage fragmentation significantly affects ECM production in vitro. Increased fragmentation enhances ECM production. CLINICAL RELEVANCE: Assuming a similar behavior in vivo, we recommend mincing the cartilage into small pieces when performing the cartilage fragment autograft implantation technique in order to increase ECM production. Further in vitro studies investigating cartilage of younger nonarthritic donors, as well as in vivo studies, are needed.
Authors: A Marmotti; S Mattia; F Castoldi; A Barbero; L Mangiavini; D E Bonasia; M Bruzzone; F Dettoni; R Scurati; G M Peretti Journal: Stem Cells Int Date: 2017-11-16 Impact factor: 5.443
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