Pelleted Bone Marrow Derived Mesenchymal Stem Cells Are Better Protected from the Deleterious Effects of Arthroscopic Heat Shock

Abstract

Introduction: 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 × 106 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.

Evaluation of in vitro chondrocytic differentiation: A stem cell research initiative at the King Abdulaziz University, Kingdom of Saudi Arabia

“…Unlike the MSCs isolated from adult tissues, the hWJSCs being relative young phenotype showed active cell proliferation with time. This was supported by earlier studies, which identified the doubling time of these hWJSCs to be much shorter [ 10 , 11 ], thus it can support generation of sufficient cell numbers needed for regenerative medicine applications.…”

“…Derived cells were analysed for the presence of MSC related CD surface markers expression using fluorescent activated cell sorting (FACS) analysis as reported earlier [ 11 ]. Briefly, independent aliquots of cells (2 x 105) were treated with MSC isotype/phenotype cocktails (Miltenyi Biotec) or combination of individual antibodies (CD29, CD44; BD Pharmingen) to avoid interference of fluorochromes.…”

Repair of Articular Cartilage Defects using Bone Marrow Derived Mesenchymal Stem Cells in Rabbits

“…Kalamegam et al Mentioned that the repair of articular cartilage injury by several methods were functionally inadequate in the long term and increased the risk of further damage [14]. Based on these reasons, the repair methods of damaged articular cartilage are limited and the traditional treatments are not ideal.…”

In vitro Evaluation of the Anti-inflammatory Effects of Thymoquinone in Osteoarthritis and in silico Analysis of Inter-Related Pathways in Age-Related Degenerative Diseases

“…BM-MSCs related CD markers expression was analyzed from cells of early passages (P1-P3) using FACS analysis as reported earlier (Kalamegam et al, 2016). Briefly, the cells were trypsinized upon reaching 70% confluence, using 0.25% Trypsin-EDTA (Life Technologies, Thermo Fisher Scientific, Waltham, MA, United States), centrifuged at 500 g × 5 min.…”