A unique porcine extracellular matrix (ECM) derived injectable tissue construct with 100 nm or 20 nm gold nanoparticles (AuNP) was developed for musculoskeletal tissue engineering applications. ECM has been shown to encourage cellularity and tissue remodeling due to its release of growth factors while AuNP have been shown to reduce reactive oxygen species (ROS) levels. Injectable tissue constructs were created by homogenizing decellularized porcine diaphragm tendon conjugated with 100 nm or 20 nm AuNP at 1x, 4x, and 8x concentrations. Extrusion force testing demonstrated that homogenized tissue constructs were injectable at an appropriate cannula size and force. L-929 murine fibroblasts were used to measure cell viability, cell proliferation, intracellular ROS levels, and cell migration in response to constructs. Enhanced cell viability and proliferation are observed on 1 × 20 nm AuNP constructs. ROS assays demonstrate reduced cellular ROS concentrations from all 20 nm AuNP constructs and from 8 × 100 nm AuNP constructs compared with constructs without nanoparticles. Cellular migration is higher towards 4 × 20 nm AuNP constructs compared with constructs without nanoparticles. Results support the potential use of a porcine ECM derived injectable tissue construct with AuNP as an injectable tissue construct to reduce inflammation and to promote tissue remodeling in musculoskeletal tissue engineering applications.
Collagen soft tissue fillers suffer from fast reabsorption, which minimizes their use as a tissue-engineered construct. Extensive cross-linking can be utilized to extend longevity, but changes in microstructure and biomechanics can have deleterious effects. To enhance longevity while still achieving a natural microstructure, gold nanoparticles (AuNPs) were conjugated to fibrilized collagen and homogenized into an injectable form for use as a soft tissue filler. A long-term animal study in Yucatan swine was conducted to assess biocompatibility and longevity. Two formulations of the AuNP-collagen were compared to porcine cross-linked collagen and commercially available hyaluronic acid (HA). The results of the study demonstrated that the AuNPs may provide enhanced longevity over 6 months compared to HA and cross-linked collagen. Irritation scores indicated that the AuNP-collagen construct (AuNP-CC) demonstrated low irritation compared to the cross-linked collagen and HA while histology scores demonstrated good biocompatibility. Overall, it may be possible to utilize AuNPs to stabilize and increase the longevity of CC while still achieving biocompatibility.
Soy lecithin has been shown to play a critical role in cell signaling and cellular membrane structure. In addition, it has been shown to increase biocompatibility, hydrophilicity, and decrease cytotoxicity. Gold nanoparticles have also shown to improve cellularity. Lecithin, gold nanoparticles, and polycaprolactone (PCL) solutions were electrospun in order to develop unique mesh materials for the treatment of osteoarthritis. The electrospinning parameters were optimized to achieve different solution ratios for fiber optimization. The amount of lecithin mixed with PCL varied from 30 wt.% to 50 wt.% . Gold nanoparticles (1% to 10% concentrations) were also added to lecithin-PCL mixture. The mechanical and chemical properties of the fiber mesh were analyzed via contact angle test, tensile mechanical tests, Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). Cell viability was measured using a WST-1 Assay. Scanning electron microscopy confirmed the successful formation of fiber mesh. The compositions of 40% soy lecithin with PCL in 40% solvent (40:40) resulted in the most well-formed fiber mesh. DSC melt temperatures were statically insignificant; uniaxial stresses and the moduli resulted in no significant difference between the test composition and pristine PCL compositions. WST-1 assay revealed all compositions were non-cytotoxic. Overall, the addition of lecithin increased hydrophilicity while maintaining cell viability and the mechanical and chemical properties of PCL. This study demonstrated that it is possible to successfully electrospin a lecithin, gold nanoparticle, and polycaprolactone scaffold for tissue engineering applications.
Surgical repair of the anterior cruciate ligament (ACL) can involve autograft or allograft materials. Allografts are typically chosen to avoid donor site morbidity associated with autografts harvest, but they can also result in a prolonged inflammatory period and delayed graft remodeling when compared to autografts. The aim of this study was to investigate the use of gold nanoparticles (AuNPs) conjugated to allografts to determine if AuNPs can reduce inflammation and enhance graft remodeling in an ovine model. Six sheep had their ACL surgically removed and replaced with a decellularized human gracilis tendon. Three of the sheep received grafts conjugated with 20 nm gold nanoparticles, while three of the sheep received grafts without the gold nanoparticles. The sheep were sacrificed 8 weeks after ACL reconstruction. Immediately following sacrifice, joint fluid was collected for cytology. Semi-quantitative histological scoring of the bone tunnel portion and the intra-articular portion of the grafts were performed independently along with descriptive analysis of histologic changes and quantitative analysis of revascularization. The results demonstrated that AuNP experimental grafts had an overall better histological scores than the non-AuNPs graft. The AuNPs grafts exhibited decreased inflammation in the bone tunnel portion of the graft, the intra-articular portion of the graft, and in the synovial fluid cell count. Overall, the results demonstrated that the grafts conjugated with nanoparticles have the potential to be influence inflammation and overall remodeling response.
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