The antimicrobial activity and biocompatibility of a controlled gentamicin-releasing single-layer sol-gel coating on hydroxyapatite-coated titanium.

AIMS: The aim of this study was to develop a single-layer hybrid organic-inorganic sol-gel coating that is capable of a controlled antibiotic release for cementless hydroxyapatite (HA)-coated titanium orthopaedic prostheses. METHODS: Coatings containing gentamicin at a concentration of 1.25% weight/volume (wt/vol), similar to that found in commercially available antibiotic-loaded bone cement, were prepared and tested in the laboratory for: kinetics of antibiotic release; activity against planktonic and biofilm bacterial cultures; biocompatibility with cultured mammalian cells; and physical bonding to the material (n = 3 in all tests). The sol-gel coatings and controls were then tested in vivo in a small animal healing model (four materials tested; n = 6 per material), and applied to the surface of commercially pure HA-coated titanium rods. RESULTS: The coating released gentamicin at > 10 × minimum inhibitory concentration (MIC) for sensitive staphylococcal strains within one hour thereby potentially giving effective prophylaxis for arthroplasty surgery, and showed > 99% elution of the antibiotic within the coating after 48 hours. There was total eradication of both planktonic bacteria and established bacterial biofilms of a panel of clinically relevant staphylococci. Mesenchymal stem cells adhered to the coated surfaces and differentiated towards osteoblasts, depositing calcium and expressing the bone marker protein, osteopontin. In the in vivo small animal bone healing model, the antibiotic sol-gel coated titanium (Ti)/HA rod led to osseointegration equivalent to that of the conventional HA-coated surface. CONCLUSION: In this study we report a new sol-gel technology that can release gentamicin from a bioceramic-coated cementless arthroplasty material. In vitro, local gentamicin levels are in excess of what can be achieved by antibiotic-loaded bone cement. In vivo, bone healing in an animal model is not impaired. This, thus, represents a biomaterial modification that may have the potential to protect at-risk patients from implant-related deep infection. Cite this article: Bone Joint J 2021;103-B(3):522-529.

In vivo safety and efficacy testing of a thermally triggered injectable hydrogel scaffold for bone regeneration and augmentation in a rat model.

Bone loss resulting from degenerative diseases and trauma is a significant clinical burden which is likely to grow exponentially with the aging population. In a number of conditions where pre-formed materials are clinically inappropriate an injectable bone forming hydrogel could be beneficial. The development of an injectable hydrogel to stimulate bone repair and regeneration would have broad clinical impact and economic benefit in a variety of orthopedic clinical applications. We have previously reported the development of a Laponite® crosslinked pNIPAM-co-DMAc (L-pNIPAM-co-DMAc) hydrogel delivery system, loaded with hydroxyapatite nanoparticles (HAPna), which was capable of inducing osteogenic differentiation of mesenchymal stem cells (MSCs) without the need for additional growth factors in vitro. However to enable progression towards clinical acceptability, biocompatibility and efficacy of the L-pNIPAM-co-DMAc hydrogel to induce bone repair in vivo must be determined. Biocompatibility was evaluated by subcutaneous implantation for 6 weeks in rats, and efficacy to augment bone repair was evaluated within a rat femur defect model for 4 weeks. No inflammatory reactions, organ toxicity or systemic toxicity were observed. In young male rats where hydrogel was injected, defect healing was less effective than sham operated controls when rat MSCs were incorporated. Enhanced bone healing was observed however, in aged exbreeder female rats where acellular hydrogel was injected, with increased deposition of collagen type I and Runx2. Integration of the hydrogel with surrounding bone was observed without the need for delivered MSCs; native cell infiltration was also seen and bone formation was observed within all hydrogel systems investigated. This hydrogel can be delivered directly into the target site, is biocompatible, promotes increased bone formation and facilitates migration of cells to promote integration with surrounding bone, for safe and efficacious bone repair.

Multilayer Nanoscale Encapsulation of Biofunctional Peptides to Enhance Bone Tissue Regeneration In Vivo.

Bone tissue healing is a dynamic process that is initiated by the recruitment of osteoprogenitor cells followed by their migration, proliferation, differentiation, and development of a mineralizing extracellular matrix. The work aims to manufacture a functionalized porous membrane that stimulates early events in bone healing for initiating a regenerative cascade. Layer-by-layer (LbL) assembly is proposed to modify the surface of osteoconductive electrospun meshes, based on poly(lactic-co-glycolic acid) and nanohydroxyapatite, by using poly(allylamine hydrochloride) and poly(sodium 4-styrenesulfonate) as polyelectrolytes. Molecular cues are incorporated by grafting peptide fragments into the discrete nanolayers. KRSR (lysine-arginine-serine-arginine) sequence is grafted to enhance cell adhesion and proliferation, NSPVNSKIPKACCVPTELSAI to guide bone marrow mesenchymal stem cells differentiation in osteoblasts, and FHRRIKA (phenylalanine-histidine-arginine-arginine-isoleucine-lysine-alanine) to improve mineralization matrix formation. Scanning electron microscopy, infrared spectroscopy, and X-ray photoelectron spectroscopy demonstrate the successful surface functionalization. Furthermore, the peptide incorporation enhances cellular processes, with good viability and significant increase of alkaline phosphatase activity, osteopontin, and osteocalcin. The functionalized membrane induces a favorable in vivo response after implantation for four weeks in nonhealing rat calvarial defect model. It is concluded that the multilayer nanoencapsulation of biofunctional peptides using LbL approach has significant potential as innovative manufacturing technique to improve bone regeneration in orthopedic and craniofacial medical devices.

Polymer-hydroxyapatite composite versus polymer interference screws in anterior cruciate ligament reconstruction in a large animal model.

The aim of the study was to assess the hard tissue response of a composite hydroxyapatite/poly L-lactic acid (HA/PLLA) interference screw for anterior cruciate ligament (ACL) reconstruction compared to a standard PLLA screw. Twelve skeletally mature rams underwent unilateral ACL reconstruction using an autologous bone-patellar tendon graft. Each animal received either two test HA/PLLA interference screws or two control PLLA interference screws. Animals were sacrificed at 6 and 12 months post-implantation and the operated knees excised. Undecalcified sections of the screw and surrounding tissues were cut from resin embedded samples and stained; sections were approximately parallel to the longitudinal axis of the screws. A quantitative assessment of bone formation between each screw type (PLLA vs. HA/PLLA) and adjacent tissue in both the tibia and femur was undertaken using automated image analysis (KS400, Zeiss, UK). The inflammatory response of each screw type was assessed by histological evaluation. New bone formation along the perimeter of the screw threads was statistically significantly higher with the HA/PLLA than the PLLA alone. The inflammatory response as assessed semi-quantitatively by histologically determining the number of inflammatory cells present in the tissue adjacent to the implant, was higher for PLLA than HA/PLLA. Significantly increased new bone formation and decreased inflammatory cells were observed in vivo with the composite screw in comparison with the standard polymer. A novel HA/PLLA composite biomaterial in the form of an interference screw demonstrated an improved hard-tissue response compared to PLLA in a large animal ACL reconstruction. This study determined the differences in the tissue response between PLLA and a composite material of HA/PLLA. The improved tissue related outcomes observed in vivo, may be of benefit clinically in ACL reconstruction.

The design and production of Co-Cr alloy implants with controlled surface topography by CAD-CAM method and their effects on osseointegration.

Improved fixation and increased longevity are still important performance criteria in the development of orthopaedic prostheses. The osseointegration of a series of implant designs made of conventional cobalt-chromium alloy was investigated, the shape of each implant being the critical variable. The shape was defined by computer-aided design with a view to maximising interdigitation of new bone with the implant. Two different process routes, conventional casting and selective laser sintering were employed, each process yielded implants that had identical surface topology but different microstructures. Hydroxyapatite (HA) was used to coat some samples by plasma spraying. Bone formation associated with each implant design was delineated through the administration of fluorescent vital dyes at three time points following their implantation into New Zealand white rabbits. After one month, specimens were harvested, resin embedded, serial sectioned and examined under fluorescent light microscopy. The amount of bone growth was quantified using image analysis. Plasma spray HA-coated samples promoted better osteogenesis and integration than uncoated samples. The extent of bone growth associated with identically shaped specimens fabricated by the SLS route was markedly greater, attributed to the microstructure of these implants.