Previous in vitro work demonstrated porous PLA and PLGA both had the mechanical strength and sustained the excellent skeletal stem cell (SSC) growth required of an osteogenic bonegraft substitute, for use in impaction bone grafting. The purpose of this investigation was to assess the effects of the addition of hydroxyapatite (HA) to the scaffolds before clinical translation. PLA, PLA+10% HA, PLGA, and PLGA+10% HA were milled and impacted into discs before undergoing a standardized shear test. Cellular compatibility analysis followed 14 days incubation with human skeletal stems cells (SSC). The best two performing polymers were taken forward for in vivo analysis. SSC seeded polymer discs were implanted subcutaneously in mice. All polymers had superior mechanical shear strength compared with allograft (p < 0.01). Excellent SSC survival was demonstrated on all polymers, but the PLA polymers showed enhanced osteoblastic activity (ALP assay p < 0.01) and collagen-1 formation. In vivo analysis was performed on PLA and PLA+10% HA. MicroCT analysis revealed increased bone formation on the PLA HA (p < 0.01), and excellent neo-vessel formation in both samples. Histology confirmed evidence of de novo bone formation. PLA HA showed both enhanced osteoinductive and osteogenic capacity. This polymer composite has been selected for scaled-up experimentation before clinical translation.

A series of random copolymers were synthesized via the copolymerization of a carbohydrate lactone, acetic acid 5-acetoxy-6-oxotetrahydropyran-2-yl methyl ester (1), and ε-caprolactone. The copolymers were characterized by nuclear magnetic resonance (NMR) spectroscopy, size exclusion chromatography (SEC), differential scanning calorimetry (DSC), and thermal gravimetric analysis (TGA). Copolymers (P1–P4) were produced with typical carbohydrate ester compositions of 1–4 mol %. These copolymers are semicrystalline and can be processed into thin films with Young’s moduli of 300–420 MPa, values that exceed that for polycaprolactone (PCL). The copolymers were processed using supercritical carbon dioxide (scCO2, 35 °C, 200 bar) into foamed, porous scaffolds, which were characterized by dynamic mechanical thermal analyses (DMTA), mercury porosimetry, and scanning electron microscopy (SEM). The copolymer foams showed increased pore size with increasing carbohydrate ester content. The average pore size increased from 71 μm (PCL) to 319 μm (P4). The foamed scaffolds have normalized storage moduli ranging from 37 MPa cm3 g–1 (P4) to 109 MPa cm3 g–1 (P1). A representative copolymer foamed scaffold, tested according to ISO 10993-5 criteria, was cytocompatible for cell culture. MC3T3 cells cultured on a film of this copolymer showed increased relative metabolic activities compared to cells cultured on a PCL film. When primary bovine chondrocytes were cultured on the foamed scaffolds, increased cell penetration into the random copolymer foam was observed compared to PCL foams.

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The development of an osteogenic bone graft substitute has important practical and cost implications in many branches of medicine where bone regeneration is required. Previous in vitro and small animal (murine) in vivo studies highlighted a porous hydroxyapatite/poly (DL-lactic acid) composite scaffold in combination with skeletal stem cells (SSCs) as a potential bone graft substitute candidate. The aim of the current study was to scale up the bone cell-scaffold construct to large animals and examine the potential for repair of a critical-sized defect via an ovine model. SSC seeded scaffolds (and unseeded scaffold controls) were implanted bilaterally into ovine femoral condyle critical defects for 3 months. A parallel in vitro analysis of ovine SSC seeded scaffolds was also performed. Post mortem mechanical indentation testing showed the bone strengths of the defect sites were 20% (controls) and 11% (SSC seeded scaffolds) those of normal cancellous bone (p < 0.01). MicroCT analysis demonstrated new bone formation within all defects with a mean increase of 13.4% in the control scaffolds over the SSC seeded scaffolds (p = 0.14). Histological examination confirmed these findings, with enhanced quality new bone within the control defects. This study highlights important issues and steps to overcome in scale-up and translation of tissue engineered products. The scaffold demonstrated encouraging results as an osteoconductive matrix; however, further work is required with cellular protocols before any human trials.

Impaction bone grafting (IBG) with human allograft remains the preferred approach for replacement of lost bone stock during revision hip surgery. Associated problems include cost, disease transmission, and stem subsidence. Synthetic grafts are therefore appealing, and ideally display similar mechanical characteristics as allograft, but with enhanced ability to form de novo bone. High and low molecular weight forms of three different polymers [poly(DL-lactide) (P(DL) LA), poly(DL-lactide-co-glycolide) (P(DL) LGA), and poly(ε-caprolactone) (PCL)] were milled, impacted into discs, and then examined in a shear testing rig, in comparison to allograft. In addition, skeletal stem cells (SSCs) were combined with each of the milled polymers, followed by impaction and examination for cell viability and number, via fluorostaining and biochemical assays. The shear strengths of high/low mwt P(DL) LA, and high/low mwt P(DL) LGA were significantly higher than allograft (p < 0.01). High/low mwt PCL had significantly lower shear strengths (p < 0.01). WST-1 assay and fluorstaining indicated significantly increased cell viability on high mwt P(DL) LA and high mwt P(DL) LGA over allograft (p < 0.05). Mechanical and biochemical analysis indicated improved properties of high mwt P(DL) LA and high mwt P(DL) LGA over allograft. This study indicates the potential of these polymers for use as substitute human allograft, creating a living composition with SSC for application in IBG.

Long hilar clamp times and anesthesia times adversely affect early postoperative renal function but not late renal function. Intraoperative bleeding adversely affected renal function at both early and late time points. Limiting intraoperative blood loss may be more important than clamp times or renal cooling in the recovery of renal function after partial nephrectomy.

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While the addition of bone cement reduced cellular viability of tissue-engineered constructs, this reduction occurred only in close proximity to the cement and measurable numbers of skeletal stem cells were observed, confirming the potential for cell population recovery.