title: Pelleted Bone Marrow Derived Mesenchymal Stem Cells Are Better Protected from the Deleterious Effects of Arthroscopic Heat Shock.
abstract: BACKGROUND
  The impact of arthroscopic temperature on joint tissues is poorly understood and it is not known how mesenchymal stem cells (MSCs) respond to the effects of heat generated by the device during the process of arthroscopy assisted experimental cell-based therapy. In the present study, we isolated and phenotypically characterized human bone marrow mesenchymal stem cells (hBMMSCs) from osteoarthritis (OA) patients, and evaluated the effect of arthroscopic heat on cells in suspension and pellet cultures.
  
  
  METHODS
  Primary cultures of hBMMSCs were isolated from bone marrow aspirates of OA patients and cultured using DMEM supplemented with 10% FBS and characterized for their stemness. hBMMSCs (1 × 10(6) cells) cultured as single cell suspensions or cell pellets were exposed to an illuminated arthroscope for 10, 20, or 30 min. This was followed by analysis of cellular proliferation and heat shock related gene expression.
  
  
  RESULTS
  hBMMSCs were viable and exhibited population doubling, short spindle morphology, MSC related CD surface markers expression and tri-lineage differentiation into adipocytes, chondrocytes and osteoblasts. Chondrogenic and osteogenic differentiation increased collagen production and alkaline phosphatase activity. Exposure of hBMMSCs to an illuminated arthroscope for 10, 20, or 30 min for 72 h decreased metabolic activity of the cells in suspensions (63.27% at 30 min) and increased metabolic activity in cell pellets (62.86% at 10 min and 68.57% at 20 min). hBMMSCs exposed to 37, 45, and 55°C for 120 s demonstrated significant upregulation of BAX, P53, Cyclin A2, Cyclin E1, TNF-α, and HSP70 in cell suspensions compared to cell pellets.
  
  
  CONCLUSIONS
  hBMMSC cell pellets are better protected from temperature alterations compared to cell suspensions. Transplantation of hBMMSCs as pellets rather than as cell suspensions to the cartilage defect site would therefore support their viability and may aid enhanced cartilage regeneration.
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz UniversityJeddah, Saudi Arabia; Sheik Salem Bin Mahfouz Scientific Chair for Treatment of Osteoarthritis by Stem Cells, King Abdulaziz UniversityJeddah, Saudi Arabia.
authors.forename: Gauthaman
authors.initials: G
authors.lastname: Kalamegam
authors.affiliation: Sheik Salem Bin Mahfouz Scientific Chair for Treatment of Osteoarthritis by Stem Cells, King Abdulaziz UniversityJeddah, Saudi Arabia; Department of Orthopedic Surgery, Faculty of Medicine, King Abdulaziz University HospitalJeddah, Saudi Arabia.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Abbas
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz UniversityJeddah, Saudi Arabia; Sheik Salem Bin Mahfouz Scientific Chair for Treatment of Osteoarthritis by Stem Cells, King Abdulaziz UniversityJeddah, Saudi Arabia; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz UniversityJeddah, Saudi Arabia.
authors.forename: Mamdooh
authors.initials: M
authors.lastname: Gari
authors.affiliation: Faculty of Applied Medical Sciences, Center of Innovation in Personalized Medicine, King Abdulaziz University Jeddah, Saudi Arabia.
authors.forename: Haneen
authors.initials: H
authors.lastname: Alsehli
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz University Jeddah, Saudi Arabia.
authors.forename: Roaa
authors.initials: R
authors.lastname: Kadam
authors.affiliation: Sheik Salem Bin Mahfouz Scientific Chair for Treatment of Osteoarthritis by Stem Cells, King Abdulaziz UniversityJeddah, Saudi Arabia; Department of Orthopedic Surgery, Faculty of Medicine, King Abdulaziz University HospitalJeddah, Saudi Arabia.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Alkaff
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz University Jeddah, Saudi Arabia.
authors.forename: Adeel
authors.initials: A
authors.lastname: Chaudhary
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz University Jeddah, Saudi Arabia.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Al-Qahtani
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz UniversityJeddah, Saudi Arabia; Faculty of Applied Medical Sciences, Center of Innovation in Personalized Medicine, King Abdulaziz UniversityJeddah, Saudi Arabia.
authors.forename: Adel
authors.initials: A
authors.lastname: Abuzenadah
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol Bristol, UK.
authors.forename: Wael
authors.initials: W
authors.lastname: Kafienah
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz UniversityJeddah, Saudi Arabia; The D-BOARD European Consortium for Biomarker Discovery, The APPROACH Innovative Medicines Initiative Consortium, Faculty of Health and Medical Sciences, University of SurreySurrey, UK; Arthritis Research UK Centre for Sport, Exercise and Osteoarthritis, Arthritis Research UK Pain Centre, Medical Research Council and Arthritis Research UK Centre for Musculoskeletal Aging Research, University of Nottingham, Queen's Medical CentreNottingham, UK.
authors.forename: Ali
authors.initials: A
authors.lastname: Mobasheri
keywords: cell viability; differentiation; fluorescence activated cell sorting (FACS); heat shock; mesenchymal stem cells; osteoarthritis (OA); population doubling