In vitro and in vivo optimization of impaction allografting by demineralization and addition of rh-OP-1

TY - JOUR

T1 - In vitro and in vivo optimization of impaction allografting by demineralization and addition of rh-OP-1

AU - Tsiridis, Eleftherios

AU - Ali, Zubier

AU - Bhalla, Amit

AU - Heliotis, Manolis

AU - Gurav, Neelam

AU - Deb, Sanjukta

AU - DiSilvio, Lucy

PY - 2007/11

Y1 - 2007/11

N2 - Impaction allografting is a bone tissue engineering technique currently used in lower limb reconstruction orthopedic surgery. Our hypothesis was that biological optimization can be achieved by demineralization and addition of osteogenic protein-1 (OP-1) to the allograft. The objective of our in vitro study was to evaluate human mesenchymal stem cell (MSC) proliferation (Alamar Blue assay, titrated thymidine assay, total DNA Hoechst 33258, and scanning electron microscopy) and osteogenic differentiation (alkaline phosphatase assay) in two types of impacted carrier, namely, demineralized bone matrix (DBM) and insoluble collagenous bone matrix (ICBM), with or without OP-1. The objective in vivo was to compare the osteogenic potential of impacted DBM with or without OP-1, with that of impacted fresh frozen allograft (FFA), again with or without OP-1. DBM + OP-1 optimized osteoinduction and significantly improved (p < 0.05) proliferation and differentiation in comparison to the majority of all other graft preparation in vitro. In addition, DBM + OP-1 was significantly superior, with regard to osteogenesis, compared to the impacted FFA alone (p < 0.001), FFA + OP-1 (p = 0.01) and DBM alone (p = 0.02) in vivo. We propose that partial demineralization and addition of OP-1 provides a good method for improving the osteoinductive properties of fresh allograft currently used in the impaction grafting technique.

AB - Impaction allografting is a bone tissue engineering technique currently used in lower limb reconstruction orthopedic surgery. Our hypothesis was that biological optimization can be achieved by demineralization and addition of osteogenic protein-1 (OP-1) to the allograft. The objective of our in vitro study was to evaluate human mesenchymal stem cell (MSC) proliferation (Alamar Blue assay, titrated thymidine assay, total DNA Hoechst 33258, and scanning electron microscopy) and osteogenic differentiation (alkaline phosphatase assay) in two types of impacted carrier, namely, demineralized bone matrix (DBM) and insoluble collagenous bone matrix (ICBM), with or without OP-1. The objective in vivo was to compare the osteogenic potential of impacted DBM with or without OP-1, with that of impacted fresh frozen allograft (FFA), again with or without OP-1. DBM + OP-1 optimized osteoinduction and significantly improved (p < 0.05) proliferation and differentiation in comparison to the majority of all other graft preparation in vitro. In addition, DBM + OP-1 was significantly superior, with regard to osteogenesis, compared to the impacted FFA alone (p < 0.001), FFA + OP-1 (p = 0.01) and DBM alone (p = 0.02) in vivo. We propose that partial demineralization and addition of OP-1 provides a good method for improving the osteoinductive properties of fresh allograft currently used in the impaction grafting technique.

KW - Adult

KW - Alkaline Phosphatase

KW - Animals

KW - Bone Demineralization Technique

KW - Bone Marrow Cells

KW - Bone Matrix

KW - Bone Morphogenetic Protein 7

KW - Bone Morphogenetic Proteins

KW - Bone Transplantation

KW - Cell Differentiation

KW - Cell Proliferation

KW - Cells, Cultured

KW - Humans

KW - Male

KW - Mesenchymal Stromal Cells

KW - Osteogenesis

KW - Rats

KW - Rats, Wistar

KW - Tissue Engineering

KW - Transplantation, Homologous

U2 - 10.1002/jor.20387

DO - 10.1002/jor.20387

M3 - Article

C2 - 17557338

SN - 0736-0266

VL - 25

SP - 1425

EP - 1437

JO - Journal of Orthopaedic Research

JF - Journal of Orthopaedic Research

IS - 11

ER -