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Expert Opinion on Biological Therapy Volume 9, 2009 - Issue 1
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Original Research

In vitro proliferation and differentiation of human mesenchymal stem cells on hydroxyapatite versus human demineralised bone matrix with and without osteogenic protein-1

Eleftherios Tsiridis Academic Unit of Trauma and Orthopaedic Surgery, Clarendon Wing, Leeds General Infirmary, Great George Street, Leeds, LS1 3EX, UK +44 113 392 2621; +44 113 392 2621; [email protected]; University College London Medical School, The Institute of Orthopaedics, Royal Free and Royal National Orthopaedic Hospital, Department of Musculoskeletal Science, Stanmore, London, HA7 4LP, UK; University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, MD MSc FRCS,
Zubier Ali University College London Medical School, The Institute of Orthopaedics, Royal Free and Royal National Orthopaedic Hospital, Department of Musculoskeletal Science, Stanmore, London, HA7 4LP, UK; University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, BSc MBBS,
Amit Bhalla University College London Medical School, The Institute of Orthopaedics, Royal Free and Royal National Orthopaedic Hospital, Department of Musculoskeletal Science, Stanmore, London, HA7 4LP, UK; University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, BSc MBBS,
Zakareya Gamie Academic Unit of Trauma and Orthopaedic Surgery, Clarendon Wing, Leeds General Infirmary, Great George Street, Leeds, LS1 3EX, UK +44 113 392 2621; +44 113 392 2621; [email protected]
, BSc MBchB,
Manolis Heliotis University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK; Norhwick Park and St' Mark's Hospital, Regional North West London Maxillofascial Unit, North West London Hospitals NHS Trust Watford Road, Harrow, London, HA1 3UJ, UK
, MBChB BDS MSc FDSRCS FRCS,
Neelam Gurav University College London Medical School, The Institute of Orthopaedics, Royal Free and Royal National Orthopaedic Hospital, Department of Musculoskeletal Science, Stanmore, London, HA7 4LP, UK; University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, PhD,
Sanjukta Deb University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, PhD &
Lucy DiSilvio University College London Medical School, The Institute of Orthopaedics, Royal Free and Royal National Orthopaedic Hospital, Department of Musculoskeletal Science, Stanmore, London, HA7 4LP, UK; University of London, Guy's Hospital, King's College London, Biomaterials and Biomimetics Group, London, SE1 9RT, UK
, PhD show all
Pages 9-19 | Published online: 08 Dec 2008
 
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Abstract

Background: Allografting is currently used in lower limb reconstruction surgery. Demineralised bone matrix (DBM) is more osteoinductive compared with allografts but lacks mechanical strength. Osteogenic protein-1 (OP-1) can improve the osteoinductivity of the allograft, however recent reports indicate significant allograft resorption when it is combined with OP-1. Objectives: Our hypothesis was that hydroxyapatite (HA) with human-mesenchymal stem cells (h-MSCs) and OP-1 (HA+h-MSCs+OP-1) has similar osteoinductive properties to human-DBM+h-MSCs. The objective was to evaluate h-MSC proliferation (by tritiated thymidine incorporation, total DNA Hoechst 33258 and scanning electron microscopy) and osteogenic differentiation (from alkaline phosphatase activity) in human demineralised bone matrix (h-DBM) and HA, with or without OP-1. Results: H-MSC proliferation on HA+OP-1 was significantly higher compared with that on HA at all time points (p < 0.05) and compared with DBM alone [day 1, (198.4 vs 95.4) p = 0.042; day 14 (286.1 vs 119.9), p < 0.001]. H-MSC proliferation was higher in DBM+OP-1, at all time points compared with HA+OP-1 but the difference was not statistically significant (p > 0.05). H-MSC differentiation was significantly higher in HA+OP-1 compared with HA (p < 0.05) but not significantly different from diffferentiation on DBM alone (p > 0.05). Differentiation was significantly higher on DBM+OP-1 at all time points compared with HA (p < 0.05) and with HA+OP-1 [day 1, (21.1 vs 10.1) (p = 0.03); day 7 (39.4 vs 7.1) (p < 0.01); day 14 (40.2 vs 14.4) (p < 0.001)]. Conclusions: When HA+h-MSCs is combined with OP-1 in vitro its osteogenic potential is similar to that of DBM+h-MSCs alone which may be adequate for non-weight-bearing applications. Mechanical testing however is of great importance for weight-bearing applications and the in vivo testing of the composite graft HA+h-MSCs+OP-1 vs DBM+h-MSCs is recommended.

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