Bone grafts prepared with selective cell retention technology heal canine segmental defects as effectively as autograft.

Using a canine critical-size segmental defect model, a two-phased study was undertaken to evaluate the healing efficacy of demineralized bone and cancellous chips (DBM-CC) enriched with osteoprogenitor cells using a Selective Cell Retention (SCR) technology. The goals of this study were: 1) to determine the bone-healing efficacy of SCR-enriched grafts versus autograft, and 2) to assess the value of clotting SCR-enriched grafts with platelet-rich plasma (PRP). Thirty dogs were included in Phase I: 18 dogs were treated with an SCR-enriched DBM-CC graft clotted with autologous bone marrow, and were compared to 12 autograft controls. In Phase II, 24 animals were divided into 4 groups of 6 animals, each treated with a different bone graft material: 1) iliac crest autograft, 2) DBM-CC alone, 3) DBM-CC saturated with marrow, and 4) SCR-enriched DBM-CC clotted with PRP. All grafts were placed unilaterally in a 21-mm long osteoperiosteal femoral, instrumented, critical-size defect. Radiographs were obtained for all animals postoperatively and every 4-16 weeks; animals were then sacrificed. All femurs were prepared for histology. Femurs in the Phase II study were also analyzed by micro-CT. At 16 weeks, healing--defined by bridging bone across the defects--was observed in 50% of the DBM-CC alone group and 67% of the DBM-CC saturated with marrow group; 100% of the autograft and SCR-enriched DBM-CC groups were healed. Histologically, grafts clotted with PRP showed more mature bone than those implanted with autologous bone, which in turn were similar to those implanted with bone marrow clotted SCR-enriched grafts. These results demonstrated that: 1) SCR-enriched DBM-CC was equivalent to autograft to repair critical-size defects, and 2) while not statistically significant, PRP may have accelerated bone maturation when used to clot osteoprogenitor-enriched DBM-CC grafts--as compared to cell-enriched, DBM-CC grafts without PRP--in large animal models.

Allogeneic mesenchymal stem cells regenerate bone in a critical-sized canine segmental defect.

BACKGROUND: Mesenchymal stem cells from adult bone marrow are multipotent cells capable of forming bone, cartilage, and other connective tissues. In a previous study, we demonstrated that autologous mesenchymal stem cells could repair a critical-sized bone defect in the dog. The objective of this study was to determine whether the use of allogeneic mesenchymal stem cells could heal a critical-sized bone defect in the femoral diaphysis in dogs without the use of immunosuppressive therapy. METHODS: A critical-sized segmental bone defect, 21 mm in length, was created in the mid-portion of the femoral diaphysis of twelve adult dogs that weighed between 22 and 25 kg. Each defect was treated with allogeneic mesenchymal stem cells loaded onto a hollow ceramic cylinder consisting of hydroxyapatite-tricalcium phosphate. A complete mismatch between donor stem cells and recipient dogs was identified by dog leukocyte antigen typing prior to implantation. The healing response was evaluated histologically and radiographically at four, eight, and sixteen weeks after implantation. The radiographic and histological results at sixteen weeks were compared with the historical data for the control defects, which included defects that had been treated with a cylinder loaded with autologous mesenchymal stem cells, defects treated with a cylinder without mesenchymal stem cells, and defects that had been left untreated (empty). The systemic immune response was evaluated by the analysis of recipient serum for production of antibodies against allogeneic cells. RESULTS: For defects treated with allogeneic mesenchymal stem cell implants, no adverse host response could be detected at any time-point. Histologically, no lymphocytic infiltration occurred and no antibodies against allogeneic cells were detected. Histologically, by eight weeks, a callus spanned the length of the defect, and lamellar bone filled the pores of the implant at the host bone-implant interface. Fluorescently labeled allogeneic cells were also detected. At sixteen weeks, new bone had formed throughout the implant. These results were consistent with those seen in implants loaded with autologous cells. Implants loaded with allogeneic or autologous stem cells had significantly greater amounts of bone within the available pore space than did cell-free implants at sixteen weeks (p < 0.05). CONCLUSIONS: The results of this study demonstrated that allogeneic mesenchymal stem cells loaded on hydroxyapatite-tricalcium phosphate implants enhanced the repair of a critical-sized segmental defect in the canine femur without the use of immunosuppressive therapy. No adverse immune response was detected in this model.

Evaluation of autologous platelet concentrate for intertransverse process lumbar fusion.

Data on the role of platelet concentrate (PC) in spinal fusion are limited. Using the New Zealand white rabbit model, we compared fusion rates at L5-L6 using 2 different volumes (1.5 cm(3), 3.0 cm(3)) of iliac crest autograft with and without PC (4 groups total, 10 animals in each). PC was collected from donor rabbits and adjusted to a concentration of 1 x 10(6) platelets/mL. Bone growth and fusion were evaluated using biomechanical, radiographic, and histologic testing. At 1.5 cm(3), autograft alone had a 29% fusion rate, compared with autograft plus PC, which had a 57% fusion rate (P = .06). At 3.0 cm(3), the fusion rate approached 90% in both groups. Radiologic fusion had a 70% correlation with biomechanical test results. Huo/Friedlaender scores were 4.3 (SD, 2.9) for 1.5-cm(3) autograft alone; 5.0 (SD, 3.5) for 1.5-cm(3) autograft plus PC; 4.7 (SD, 2.5) for 3.0-cm(3) autograft alone; and 7.7 (SD, 0.6) for 3.0-cm(3) autograft plus PC. For 1.5-cm(3) autograft, a trend toward improvement in biomechanically defined fusion was found when PC was added, which suggests that, when the amount of bone graft is limited, PC may function as a graft extender in posterolateral fusion. At higher volumes of bone graft, no appreciable difference was noted between groups. Although radiography revealed fusion masses, the technique was not useful in identifying pseudarthrosis. On histologic analysis, adding PC seemed to result in more mature bone at both volumes, with the most mature bone in the group with 3.0-cm(3) autograft plus PC.

Efficacy of mesenchymal stem cell enriched grafts in an ovine posterolateral lumbar spine model.

STUDY DESIGN: Four groups of 6 animals underwent single-level noninstrumented posterolateral lumbar fusion (PLF) with one of the following grafts: 1) autograft, 2) cell-enriched beta-tricalcium phosphate (TCP), 3) TCP with whole bone marrow, and 4) TCP alone. Plain radiographs were taken after surgery and at death, 6 months after surgery. Explanted spine segments were analyzed by manual palpation, micro-CT, and histology. OBJECTIVE: A sheep spine fusion study was undertaken to evaluate the healing performance of a TCP graft enriched with osteoprogenitor cells using Selective Cell Retention technology (SCR), compared with autograft, TCP with whole bone marrow, and TCP alone. SUMMARY OF BACKGROUND DATA: Improved bone healing with previously demonstrated using grafts enriched in osteoprogenitor cells. METHODS: Cell-enriched grafts were obtained by processing 30 mL of bone marrow through 10 mL of TCP. TCP was also used either saturated with bone marrow or alone. RESULTS: At 6 months, 33% of the SCR-enriched TCP and 25% of the autograft sites were fused, compared with 8% of the TCP plus whole bone marrow and 0% of the TCP alone. Histology of fused samples showed denser bone formation in the SCR-enriched TCP grafts than in the autograft sites. CONCLUSIONS: The use of SCR-enriched TCP and autograft resulted in similar fusion rates in an ovine posterolateral noninstrumented lumbar spine fusion model.

Investigation of allogeneic mesenchymal stem cell-based alveolar bone formation: preliminary findings.

This study was designed to evaluate mesenchymal stem cell (MSC)-based alveolar bone regeneration in a canine alveolar saddle defect model. MSCs were loaded onto hydroxyapatite/tricalcium phosphate (HA/TCP) matrices. Scanning electron microscopic (SEM) evaluation demonstrated greater than 75% MSC coverage of the HA/TCP porous surface prior to placement regardless of MSC donor. Matrices, 6 mm x 6 mm x 20 mm, with and without cells, were implanted for 4 and 9 weeks, then removed for histological evaluation of bone formation. Cell-free control matrices were compared with MSC-loaded matrices post implantation. Histomorphometrical analysis showed that equivalent amounts of new bone were formed within the pores of the matrices loaded with autologous MSCs or MSCs from an unrelated donor. Bone formation in the cell-free HA/TCP matrices was less extensive. There was no histologic evidence of an immunological response to autologous MSCs. Surprisingly, allogeneic MSC implantation also failed to provoke an immune response. Analysis of circulating antibody levels against MSCs supported the hypothesis that neither autologous nor allogeneic MSCs induced a systemic response by the host. Analysis of dye-labelled MSCs in histological sections confirmed that the MSCs persisted in the implants throughout the course of the experiment. At 9 weeks, labelled cells were present within the lacunae of newly formed bone. We conclude that autologous and allogeneic MSCs have the capacity to regenerate bone within craniofacial defects.