Measurement of migration of a humeral head resurfacing prosthesis using radiostereometry without implant marking: An experimental study

Today, the shoulder joint is the third most commonly replaced joint after the hip and knee joints and the incidence is increasing. In Sweden, 1863 primary Shoulder Arthroplasties and 195 revisions were performed in 2017. The most common diagnoses are Osteoarthritis and irreparable tears of the rotator cuff, with or without arthropathy, often referred to as cuff tear arthropathy. Different Shoulder Arthroplasty (SA) concepts include anatomical total shoulder arthroplasty (TSA), hemiarthroplasty (HSA) and reversed shoulder arthroplasty, but also humeral head resurfacing (HHR) and stemless arthroplasties. All concepts offer pain relief, improvement of function and in quality of life for the different diagnoses. Unfortunately, there are sometimes complications after SA. They involve periprosthetic joint infection, humeral and glenoid fractures, stress shielding, loosening of the glenoid and humeral component but also glenoid erosion and cuff rupture. Some of these complications are most common within 1 year after operation, some after several years, both may lead to a revision. This, together with the fact that new designs of implants and methods of fixation of SA continues to develop, stresses the importance of continuous monitoring of implant survival and follow-up. The overall aim of this thesis was to describe clinical examples of different methods to assess the outcome after Shoulder Arthroplasty. The most common methods are clinical examination, radiographic assessment, Patient Reported Outcome Measure (PROM), National Joint registries, where revisions are an important outcome, but also Clinical Trials. All of these methods are used in one or more of the 4 papers in this thesis and shows the complexity of the topic and the practical work. In paper I we used Radio Stereometric Analysis (RSA) in an experimental set-up and concluded that marker-free RSA can be used for a humeral head resurfacing arthroplasty. In paper II we used data from the Swedish Shoulder Arthroplasty Registry (SSAR) with PROM and revisions to conclude that age is the only factor that affects revision when comparing HSA and HHR. Paper III is a long-time follow-up of a Randomized controlled study where we used radiological assessment, PROM and revisions. The conclusion was that both TSA and HSA develop severe radiological changes 10 year after primary operation. Paper IV is a prospective RSA cohort study where we also evaluated PROM and revisions. The conclusion is that HHR seems to obtain a secure fixation in the humerus, after an initial migration. But also that the prostheses shows continuous glenoid wear. The main conclusion of this thesis is that patient’s operated with SA needs continuous monitoring and several methods may be used to evaluate the outcome

Experimental mapping of soluble protein domains using a hierarchical approach

Exploring the function and 3D space of large multidomain protein targets often requires sophisticated experimentation to obtain the targets in a form suitable for structure determination. Screening methods capable of selecting well-expressed, soluble fragments from DNA libraries exist, but require the use of automation to maximize chances of picking a few good candidates. Here, we describe the use of an insertion dihydrofolate reductase (DHFR) vector to select in-frame fragments and a split-GFP assay technology to filter-out constructs that express insoluble protein fragments. With the incorporation of an IPCR step to create high density, focused sublibraries of fragments, this cost-effective method can be performed manually with no a priori knowledge of domain boundaries while permitting single amino acid resolution boundary mapping. We used it on the well-characterized p85α subunit of the phosphoinositide-3-kinase to demonstrate the robustness and efficiency of our methodology. We then successfully tested it onto the polyketide synthase PpsC from Mycobacterium tuberculosis, a potential drug target involved in the biosynthesis of complex lipids in the cell envelope. X-ray quality crystals from the acyl-transferase (AT), dehydratase (DH) and enoyl-reductase (ER) domains have been obtained

Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

To facilitate fine-scale phenotyping of whole specimens, we describe here a set of tissue fixation-embedding, detergent-clearing and staining protocols that can be used to transform excised organs and whole organisms into optically transparent samples within 1–2 weeks without compromising their cellular architecture or endogenous fluorescence. PACT (passive CLARITY technique) and PARS (perfusion-assisted agent release in situ) use tissue-hydrogel hybrids to stabilize tissue biomolecules during selective lipid extraction, resulting in enhanced clearing efficiency and sample integrity. Furthermore, the macromolecule permeability of PACT- and PARS-processed tissue hybrids supports the diffusion of immunolabels throughout intact tissue, whereas RIMS (refractive index matching solution) grants high-resolution imaging at depth by further reducing light scattering in cleared and uncleared samples alike. These methods are adaptable to difficult-to-image tissues, such as bone (PACT-deCAL), and to magnified single-cell visualization (ePACT). Together, these protocols and solutions enable phenotyping of subcellular components and tracing cellular connectivity in intact biological networks

Markerless Roentgen Stereophotogrammetric Analysis for in vivo implant migration measurement using three dimensional surface models to represent bone

Optimising joint reconstruction management in arthritis and bone tumour patient