Mesenchymal stem cells (MSC) are attractive candidates for bone regeneration approaches. Benefits of MSC therapy are mainly attributed to paracrine effects via soluble factors, exerting both immunoregulatory and regenerative actions. Encapsulation of MSC in hydrogels prepared with extracellular matrix (ECM) proteins has been proposed as a strategy to enhance their survival and potentiate their function after implantation. Functional activity of MSC can be regulated by the physical and mechanical properties of their microenvironment. In this work, we investigated whether matrix stiffness can modulate the crosstalk between MSC encapsulated in collagen hydrogels with macrophages and osteoblasts. Collagen hydrogels with a final collagen concentration of 1.5, 3 and 6 mg/mL loaded with human MSC were prepared. Viscoelastic properties of hydrogels were measured in a controlled stress rheometer. Cell distribution into the hydrogels was examined using confocal microscopy and the levels of the immunomodulatory factors interleukin-6 (IL-6) and prostaglandin E2 (PGE2) released by MSC were quantified by immunoassays. To determine the effect of matrix stiffness on the immunomodulatory potential of MSC, human macrophages obtained from healthy blood were cultured in media conditioned by MSC in hydrogels. The involvement of IL-6 and PGE2 in MSC-mediated immunomodulation was investigated employing neutralizing antibodies. Finally, the influence of soluble factors released by MSC in hydrogels on bone-forming cells was studied using osteoblasts obtained from trabecular bone explants from patients with osteonecrosis of the femoral head during total hip arthroplasty.Introduction and Objective
Materials and Method
Acrylic bone cements are used rather extensively in orthopedic and spinal applications. The incorporation of calcium phosphate additives to bone cements, to induce osteoconductivity, have typically resulted in increased cement viscosity, decreased handling, and detrimental effects of the mechanical performance of the cement. Additionally, bioactive bone cements are offered at a premium cost, which limits clinical use of these materials. The goal of this study was to examine and characterize an alternative two-solution poly (methyl Methacrylate) (PMMA) bone cement (referred to as TSBC), after incorporation of several calcium phosphate additives and antimicrobials. These bioactive and antimicrobial two-solution cements were designed to have adjustable properties that meet specific requirements of orthopedic applications. The addition of a bioactive agent would lead to increased levels of bone reformation after surgery, while an antibiotic within the cement would decrease the ability for pathogens to grow in the interface between the bone and new implant. TSBC is a pre-mixed bone cement that exhibits a combination of attractive properties including high strength, adjustable viscosity, adequate exothermal properties, as well as offering the possibility of using the same batch multiple times. The addition of antibiotics has not been previously explored in two-solution bone cements. Therefore, it is desirable to induce antibacterial activity with this formulation. Hydroxyapatite (Ca5(PO4)3(OH)), Brushite (CaHPO4•2H2O), and Tricalcium Phosphate (Ca3(PO4)2)(TCP) were incorporated into the TSBC in varying concentrations (25 and 50 wt%), and the rheological characteristics were examined to verify the feasibility of adding high concentrations of fillers to this cement formulation. Results demonstrated that unlike commercial powder-liquid formulations, calcium phosphate additives in TSBC do not detrimentally affect handling and the rheological properties of the material, while also providing maintenance of cement strength and other physical properties. TSBC material spends a dramatically increased amount of time in the swelling phase, as compared to powder-liquid formulations and thus is better suited to incorporate additives fully into its polymer matrix. Current two-solution bone cements do not contain any osteoconductive or antimicrobial agents. This study investigated the effects of addition of these bioactive agents in the physical and mechanical properties of the cement. Cement porosity was investigated to ensure that the porous nature of the bioactive cement does not damage the mechanical stability of the material. Further imaging will be conducted to demonstrate the improved osteointegration of these bioactive cement with osteoblasts (Figure 1). Degradation studies have been conducted to validate the biodegradable properties of the bioactive components and antibiotics release profile. It is further hypothesized that the degradation time will correlate to the antimicrobial activity. As the cement is replaced with natural bone, more and more antimicrobial will become exposed to the physiologic environment causing a continuous antimicrobial release as the material is partially replaced with new bone over time. Antimicrobial effectiveness and antimicrobial release studies are under-way to illustrate the cements ability to restrict growth at the cement surface, as well as show the antimicrobial release profile over time.
Many studies have looked at the effect of titanium versus cobalt chrome baseplates on backside wear. However, the surface finish of the materials is usually different [1,2]. There may also be subtle locking mechanism design changes [2]. The purpose of this study was to evaluate the wear performance of polyethylene inserts when mated with titanium baseplates to cobalt chrome baseplates, where both have non-polished topside surfaces and an identical locking mechanism. A total of three trays per material were used. The titanium trays are intended for cementless application and include a porous titanium surface on the underside, while the cobalt chrome trays are intended for cemented applications. All trays were Triathlon design (Stryker Orthopaedics, Mahwah, NJ). Tibial inserts were manufactured from GUR 1020 polyethylene then vacuum/flush packaged and sterilized in nitrogen (30 kGy). Cobalt chrome femoral components were articulated against the tibial inserts. Surface roughness of the baseplates was measured prior to testing using white light interferometry (Zygo, Middlefield, CT). A 6-station knee simulator (MTS, Eden Prairie, MN) was used for testing. A normal walking profile was applied [3]. Testing was conducted for 1 million cycles. A lubricant of Alpha Calf Fraction serum (Hyclone Labs, Logan, UT) diluted to 50% with a pH-balanced 20-mMole solution of deionized water and EDTA was used [4]. The serum solution was replaced and inserts were weighed for wear every 0.5 million cycles. Standard test protocols were used for cleaning, weighing, and assessing the wear loss [5]. Soak control specimens were used to correct for fluid absorption. Statistical analysis was performed using the Student's t-test (p < 0.05).INTRODUCTION
MATERIALS AND METHODS:
