Leonardo do Nascimento1, Natália Fontana Nicoletti2, Manuela Peletti-Figueiró2, Daniel Marinowic3, Asdrubal Falavigna1,2,4. 1. Health Sciences Graduate Program, Universidade de Caxias do Sul, Caxias do Sul, Brazil. 2. Cell Therapy Laboratory, Universidade de Caxias do Sul, Caxias do Sul, Brazil. 3. Brain Institute of Rio Grande do Sul, Graduate Program in Medicine and Health Sciences and School of Medicine, Pontifical Catholic University of Rio Grande do Sul, Porto Alegre, Brazil. 4. Laboratory of Basic Studies on Spinal Cord Pathologies, Department of Neurosurgery, University of Caxias of Sul, Brazil.
Abstract
OBJECTIVES: This study aimed to evaluate the efficacy of hyaluronic acid in the viability and proliferation profile of human femoral-tibial joint cartilage affected by osteoarthritis using in vitro models of chondrocytes in a 2-dimensional (2D)- and 3-dimensional (3D)-based culture model by spheroids. DESIGN: In vitro study of knee cartilage affected by osteoarthritis that required surgical treatment. Samples were cultured and exposed to hyaluronic acid (100 and 500 μM; intervention group) or vehicle solution. In monolayer or 2D culture, proliferation and cell viability were measured, and nuclear morphometry was analyzed by 4',6'-diamino-2-fenil-indol (DAPI) staining. The 3D-based culture established from the culture of articular cartilage of patients submitted to total knee arthroplasty evaluated the diameter, viability, and fusion ability of the chondrospheres created. RESULTS: Samples from 3 patients resulted in viable cultures, with chondrocyte cells exhibiting a potential for cell proliferation and viability to establish a culture. Hyaluronic acid (100 and 500 μM) improved chondrocyte viability and proliferation up to 72 hours in contact when compared with the control group, and no nuclear irregularities in morphology cell characteristics were observed by DAPI. In the 3D evaluation, hyaluronic acid (500 μM) improved the cellular feedback mechanisms, increasing the survival and maintenance of the chondrospheres after 7 days of analysis, showing the intrinsic capacity of chondrospheres grouped in the attempt to rearrange and reestablish new articular tissue. CONCLUSIONS: The 2D- and 3D-based culture models with hyaluronic acid improved chondrocyte viability and proliferation and demonstrated the ability of freshly formed chondrospheres to undergo fusion when placed together in the presence of hyaluronic acid.
OBJECTIVES: This study aimed to evaluate the efficacy of hyaluronic acid in the viability and proliferation profile of human femoral-tibial joint cartilage affected by osteoarthritis using in vitro models of chondrocytes in a 2-dimensional (2D)- and 3-dimensional (3D)-based culture model by spheroids. DESIGN: In vitro study of knee cartilage affected by osteoarthritis that required surgical treatment. Samples were cultured and exposed to hyaluronic acid (100 and 500 μM; intervention group) or vehicle solution. In monolayer or 2D culture, proliferation and cell viability were measured, and nuclear morphometry was analyzed by 4',6'-diamino-2-fenil-indol (DAPI) staining. The 3D-based culture established from the culture of articular cartilage of patients submitted to total knee arthroplasty evaluated the diameter, viability, and fusion ability of the chondrospheres created. RESULTS: Samples from 3 patients resulted in viable cultures, with chondrocyte cells exhibiting a potential for cell proliferation and viability to establish a culture. Hyaluronic acid (100 and 500 μM) improved chondrocyte viability and proliferation up to 72 hours in contact when compared with the control group, and no nuclear irregularities in morphology cell characteristics were observed by DAPI. In the 3D evaluation, hyaluronic acid (500 μM) improved the cellular feedback mechanisms, increasing the survival and maintenance of the chondrospheres after 7 days of analysis, showing the intrinsic capacity of chondrospheres grouped in the attempt to rearrange and reestablish new articular tissue. CONCLUSIONS: The 2D- and 3D-based culture models with hyaluronic acid improved chondrocyte viability and proliferation and demonstrated the ability of freshly formed chondrospheres to undergo fusion when placed together in the presence of hyaluronic acid.
Authors: Nikolai P Omelyanenko; Pavel A Karalkin; Elena A Bulanova; Elizaveta V Koudan; Vladislav A Parfenov; Sergei A Rodionov; Alisa D Knyazeva; Vladimir A Kasyanov; Igor I Babichenko; Tamara Z Chkadua; Yusef D Khesuani; Anna A Gryadunova; Vladimir A Mironov Journal: Cartilage Date: 2018-09-15 Impact factor: 4.634
Authors: Marijn Rutgers; Laura B Creemers; Kiem Gie Auw Yang; Natasja J H Raijmakers; Wouter J A Dhert; Daniel B F Saris Journal: Acta Orthop Date: 2014-08-20 Impact factor: 3.717