Analysis of the Effects of Five Factors Relevant to In Vitro Chondrogenesis of Human Mesenchymal Stem Cells Using Factorial Design and High Throughput mRNA-Profiling

The in vitro process of chondrogenic differentiation of mesenchymal stem cells for tissue engineering has been shown to require three-dimensional culture along with the addition of differentiation factors to the culture medium. In general, this leads to a phenotype lacking some of the cardinal features of native articular chondrocytes and their extracellular matrix. The factors used vary, but regularly include members of the transforming growth factor β superfamily and dexamethasone, sometimes in conjunction with fibroblast growth factor 2 and insulin-like growth factor 1, however the use of soluble factors to induce chondrogenesis has largely been studied on a single factor basis. In the present study we combined a factorial quality-by-design experiment with high-throughput mRNA profiling of a customized chondrogenesis related gene set as a tool to study in vitro chondrogenesis of human bone marrow derived mesenchymal stem cells in alginate. 48 different conditions of transforming growth factor β 1, 2 and 3, bone morphogenetic protein 2, 4 and 6, dexamethasone, insulin-like growth factor 1, fibroblast growth factor 2 and cell seeding density were included in the experiment. The analysis revealed that the best of the tested differentiation cocktails included transforming growth factor β 1 and dexamethasone. Dexamethasone acted in synergy with transforming growth factor β 1 by increasing many chondrogenic markers while directly downregulating expression of the pro-osteogenic gene osteocalcin. However, all factors beneficial to the expression of desirable hyaline cartilage markers also induced undesirable molecules, indicating that perfect chondrogenic differentiation is not achievable with the current differentiation protocols

Morbidly Obese Patients—Who Undergoes Bariatric Surgery?

Treatment seeking patients with severe obesity might choose between specialized medical treatment and surgical treatment. Knowledge of what distinguishes patients that choose either treatment is sparse, with greater understanding also needed on what consequences this choice has for the prevalence, remission and new onset of comorbidities, as well as for the bioavailability of drugs. This has prompted the studies in Gunn Signe Jakobsen and her coauthors work on treatment seeking patients with severe obesity focusing on the prevalence of comorbidities, changes in the use of drugs for hypertension, diabetes and dyslipidaemia, as well as changes in bioavailability of atorvastatin. The methods used in the studies in the thesis; "Bariatric surgery and specialized medical treatment for severe obesity Impact on cardiovascular risk factors and postsurgical pharmacokinetics of atorvastatin "; are a cross-sectional study, a registry based cohort study and a prospective pharmacokinetic study. The results of the studies presented were: - The type and number of comorbidities associated with morbid obesity did not necessarily impact upon choice of treatment, but there was an increased odds for choosing surgery for patients with higher BMI, younger age and earlier onset of obesity. - Patients opting for bariatric surgery as opposed to specialized medical treatment had higher odds of experiencing remission, and significantly lower odds for new-onset of drug treated hypertension, diabetes and dyslipidaemia. Bariatric surgery seemed to not only induce remission but was also effective in preventing disease. - The bioavailability of atorvastatin was increased after bariatric surgery, with a normalization in the long term. This knowledge can give a better understanding of the population of patients seeking treatment for severe obesity and should be included in the shared decision process when helping the patient identify their preferences for treatment of severe obesity in the context of their values

A preliminary simulation-based qualitative study of healthcare students’ experiences of interprofessional primary care scenarios

Background Introducing interprofessional education (IPE) in healthcare curricula can prepare students for healthcare practices that have become increasingly complex. The use of simulation is promoted to support IPE. This study explores healthcare students’ experiences of participating in common, sub-acute patient scenarios that routinely occur in clinical practice in primary care. More specifically, it looks at how sub-acute patient scenarios from primary care can help develop interprofessional collaborative competence. Methods Medical students (N = 10), master’s students in advanced geriatric nursing (N = 8) and bachelor’s students in nursing (N = 9) participated in the simulations. The students were in their last or second-to-last year of education. We conducted five semi-structured focus group interviews with the participants’ directly after the simulation training to elicit experiences related to the scenarios, the simulation and interprofessional collaboration. The transcripts were analysed using systematic text condensation. To supplement the focus group interviews, the students also completed the interprofessional collaborative competency attainment survey (ICCAS), which measures the students’ self-assessed interprofessional competence. Results Three main themes emerged from the analysis of the focus group interviews: realism, uncertainty and reflection. The students emphasised the importance of authentic and recognisable scenarios. They said the vague and unspecific patient symptoms created uncertainty in the situation, making it difficult to understand the patient’s diagnosis. Despite that uncertainty, they described the experience as positive. Further, the students expressed that the simulation increased their confidence in interprofessional collaboration and prepared them for future work. The results from the ICCAS questionnaire showed that the students reported a subjective positive change in their interprofessional competence after participating in the scenarios. Conclusions This study showed that simulation-based IPE with sub-acute primary care scenarios contributes to develop interprofessional collaborative competence in healthcare education. Sub-acute scenarios can supplement the more common approaches with acute care scenarios and aid in developing the collaborative competence required to work in healthcare teams

Bone marrow mesenchymal stem cells in a hyaluronan scaffold for treatment of an osteochondral defect in a rabbit model

The purpose of this study was to evaluate the efficiency of using mesenchymal stem cells (MSC) in a hyaluronan scaffold for repair of an osteochondral defect in rabbit knee. Bone marrow was harvested from the posterior iliac crest in 11 New Zealand White rabbits. MSC were isolated and cultured in autologous serum for 28 days and transferred to a hyaluronan scaffold 24 h prior to implantation. A 4 mm diameter and 1.5 mm deep defect was created in the medial femoral condyle of both knees and the scaffold with MSC was implanted in one knee while an empty scaffold was implanted in the contra-lateral knee. After 24 weeks the rabbits were killed and histological sections were subjected to semiquantitative and quantitative evaluation by observers blinded regarding treatment modality. High degree of filling was obtained, but there was no statistically significant difference between the two treatments. However, there was a tendency for a better quality of repair in the MSC treated knees. No hypertrophy was observed by either method. MSC in a hyaluronan scaffold may be a promising treatment approach, but further studies are needed to determine the best combination of scaffold and cells

Statistical analysis of main effects and interactions at day 7.

<p><b>A.</b> Normal plot of the standardized effects with the response set to mean expression of wanted markers. <b>B.</b> Corresponding main effects plot of all factors. <b>C</b> and <b>D.</b> Corresponding plots of significant second order interactions. <b>E.</b> Normal plot of the standardized effects with the response set to mean expression of unwanted markers. <b>F.</b> Corresponding main effects plot of all factors. <b>G</b> and <b>H</b>. Corresponding plots of significant second order interactions.</p