Initial studies. For the first initial study, the experimental design featured one treatment group with two mechanical loading conditions. Defects received morselized autograft prepared by mincing the excised cortical biopsy in sterile phosphate-buffered saline (PBS) [Autograft] contained within an electrospun, perforated poly-(ε-caprolactone) (PCL) nanofiber mesh tube. Limbs were stabilized with stiff [Stiff] or axially compliant [Compliant] fixation plates initially implanted in a locked configuration to prevent loading (kaxial = 250 ± 35 N/mm), but after 4 weeks, the plates were surgically unlocked to enable load transfer (kaxial = 8.0 ± 3.5 N/mm) (n = 6 to 8 per group) (4345). For the second initial study, the experimental design featured three treatment groups with one mechanical loading condition. Defects received (i) mesenchymal condensations composed of three microparticle-containing hMSC sheets for a total of 6.0 × 106 cells with BMP-2–loaded MCM (1.9 μg) and unloaded GM [BMP-2 (hMSCs)], (ii) BMP-2 (1.9 μg) soaked onto 8-mm precut absorbable collagen sponge (INFUSE Bone Graft, Medtronic, Memphis, TN) 15 min before implantation [BMP-2 (collagen)], or (iii) morselized autograft in sterile PBS [Autograft], each contained within an electrospun, perforated PCL nanofiber mesh tube. Limbs were stabilized with stiff fixation plates [Stiff] that limit load transfer (kaxial = 260 ± 28 N/mm) (n = 7 to 10 per group), modified from prior studies (4345).

Main study. The experimental design featured three treatment groups with two mechanical loading conditions (Table 1). Defects received mesenchymal condensations composed of three microparticle-containing hMSC sheets for a total of 6.0 × 106 cells incorporated with (i) unloaded MCM and GM [Empty/Control], (ii) BMP-2–loaded MCM (1.9 μg) and unloaded GM [BMP-2], or (iii) BMP-2–loaded MCM (1.9 μg) and TGF-β1–loaded GM (1.8 μg) [BMP-2 + TGF-β1] contained within an electrospun, perforated PCL nanofiber mesh tube. Limbs were stabilized with stiff [Stiff] or axially compliant [Compliant] fixation plates initially implanted in a locked configuration to prevent loading (kaxial = 250 ± 35 N/mm), but after 4 weeks, the plates were surgically unlocked to enable load transfer (kaxial = 8.0 ± 3.5 N/mm) (n = 3 to 10 per group) (4345).

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