Endothelial cells influence the osteogenic potential of bone marrow stromal cells

<p>Abstract</p> <p>Background</p> <p>Improved understanding of the interactions between bone cells and endothelial cells involved in osteogenesis should aid the development of new strategies for bone tissue engineering. The aim of the present study was to determine whether direct communication between bone marrow stromal cells (MSC) and human umbilical vein endothelial cells (EC) could influence the osteogenic potential of MSC in osteogenic factor-free medium.</p> <p>Methods</p> <p>After adding EC to MSC in a direct-contact system, cell viability and morphology were investigated with the WST assay and immnostaining. The effects on osteogenic differentiation of adding EC to MSC was systematically tested by the using Superarray assay and results were confirmed with real-time PCR.</p> <p>Results</p> <p>Five days after the addition of EC to MSC in a ratio of 1:5 (EC/MSC) significant increases in cell proliferation and cellular bridges between the two cell types were detected, as well as increased mRNA expression of alkaline phosphatase (ALP). This effect was greater than that seen with addition of osteogenic factors such as dexamethasone, ascorbic acid and β-glycerophosphate to the culture medium. The expression of transcription factor Runx2 was enhanced in MSC incubated with osteogenic stimulatory medium, but was not influenced by induction with EC. The expression of Collagen type I was not influenced by EC but the cells grown in the osteogenic factor-free medium exhibited higher expression than those cultured with osteogenic stimulatory medium.</p> <p>Conclusion</p> <p>These results show that co-culturing of EC and MSC for 5 days influences osteogenic differentiation of MSC, an effect that might be independent of Runx2, and enhances the production of ALP by MSC.</p

Temporomandibular joint pain and associated magnetic resonance findings: a retrospective study with a control group

Background To better understand and evaluate clinical usefulness of magnetic resonance imaging (MRI) in diagnosis and treatment of temporomandibular disorders (TMD), parameters for the evaluation are useful. Purpose To assess a clinically suitable staging system for evaluation of MRI of the temporomandibular joint (TMJ) and correlate the findings with age and some clinical symptoms of the TMJ. Material and Methods Retrospective analysis of 79 consecutive patients with clinical temporomandibular disorder or diagnosed inflammatory arthritis. Twenty-six healthy volunteers were included as controls. Existing data included TMJ pain, limited mouth opening (<30 mm) and corresponding MRI evaluations of the TMJs. Results The patients with clinical TMD complaints had statistically significantly more anterior disc displacement (ADD), disc deformation, caput flattening, surface destructions, osteophytes, and caput edema diagnosed by MRI compared to the controls. Among the arthritis patients, ADD, effusion, caput flattening, surface destructions, osteophytes, and caput edema were significantly more prevalent compared to the healthy volunteers. In the control group, disc deformation and presence of osteophytes significantly increased with age, and a borderline significance was found for ADD and surface destructions on the condylar head. No statistically significant associations were found between investigated clinical and MRI parameters. Conclusion This study presents a clinically suitable staging system for comparable MRI findings in the TMJs. Our results indicate that some findings are due to age-related degenerative changes rather than pathological changes. Results also show that clinical findings such as pain and limited mouth opening may not be related to changes diagnosed by MRI.publishedVersio

Translation of a standardized manufacturing protocol for mesenchymal stromal cells: A systematic comparison of validation and manufacturing data

Background Many data are available on expansion protocols for mesenchymal stromal cells (MSCs) for both experimental settings and manufacturing for clinical trials. However, there is a lack of information on translation of established protocols for Good Manufacturing Practice (GMP) from validation to manufacturing for clinical application. We present the validation and translation of a standardized pre-clinical protocol for isolation and expansion of MSCs for a clinical trial for reconstitution of alveolar bone. Methods Key parameters of 22 large-scale expansions of MSCs from bone marrow (BM) for validation were compared with 11 expansions manufactured for the clinical trial “Jaw bone reconstruction using a combination of autologous mesenchymal stromal cells and biomaterial prior to dental implant placement (MAXILLO1)” aimed at reconstruction of alveolar bone. Results Despite variations of the starting material, the robust protocol led to stable performance characteristics of expanded MSCs. Manufacturing of the autologous advanced therapy medicinal product MAXILLO-1-MSC was possible, requiring 21 days for each product. Transport of BM aspirates and MSCs within 24 h was guaranteed. MSCs fulfilled quality criteria requested by the national competent authority. In one case, the delivered MSCs developed a mosaic in chromosomal finding, showing no abnormality in differentiation capacity, growth behavior or surface marker expression during long-term culture. The proportion of cells with the mosaic decreased in long-term culture and cells stopped growth after 38.4 population doublings. Conclusions Clinical use of freshly prepared MSCs, manufactured according to a standardized and validated protocol, is feasible for bone regeneration, even if there was a long local distance between manufacturing center and clinical site. Several parameters, such as colony forming units fibroblasts (CFU-F), percentage of CD34+ cells, cell count of mononuclear cells (MNCs) and white blood cells (WBCs), of the BM may serve as a predictive tool for the yield of MSCs and may help to avoid unnecessary costs for MSC manufacturing due to insufficient cell expansion rates.publishedVersio

Peripheral odontogenic keratocyst causing bone resorption: Report of two cases

Two cases of peripheral odontogenic keratocyst (POKC) of the gingiva presenting as asymptomatic nodules are described. The exceptional finding of cystic radiolucency in POKCs was observed on dental radiography in one case, which had a diameter of 10 mm. Conservative surgical excision was performed on both cases, and histopathological examination showed features consistent with keratocysts. POKC may recur and should thus be followed up like intraosseous keracysts. No recurrences were observed after 10-years in our two cases. POKC is managed by conservative treatment. Recurrences should be excised more radically. Adjunct treatment with modified Carnoy's solution or topical 5Fluorouracil should be considered. POKC should be included in the differential diagnosis of asymptomatic gingival nodules

Efficacy of treating segmental bone defects through endochondral ossification: 3D printed designs and bone metabolic activities

Three-dimensional printing (3D printing) is a promising technique for producing scaffolds for bone tissue engineering applications. Porous scaffolds can be printed directly, and the design, shape and porosity can be controlled. 3D synthetic biodegradable polymeric scaffolds intended for in situ bone regeneration must meet stringent criteria, primarily appropriate mechanical properties, good 3D design, adequate biocompatibility and the ability to enhance bone formation. In this study, healing of critical-sized (5 ​mm) femur defects of rats was enhanced by implanting two different designs of 3D printed poly(l-lactide-co-ε-caprolactone) (poly(LA-co-CL)) scaffolds seeded with rat bone marrow mesenchymal stem cells (rBMSC), which had been pre-differentiated in vitro into cartilage-forming chondrocytes. Depending on the design, the scaffolds had an interconnected porous structure of 300–500 ​μm and porosity of 50–65%. According to a computational simulation, the internal force distribution was consistent with scaffold designs and comparable between the two designs. Moreover, the defects treated with 3D-printed scaffolds seeded with chondrocyte-like cells exhibited significantly increased bone formation up to 15 weeks compared with empty defects. In all experimental animals, bone metabolic activity was monitored by positron emission tomography 1, 3, 5, 7, 11 and 14 weeks after surgery. This demonstrated a time-dependent relationship between scaffold design and metabolic activity. This confirmed that successful regeneration was highly reproducible. The in vitro and in vivo data indicated that the experimental setups had promising outcomes and could facilitate new bone formation through endochondral ossification.publishedVersio

Temporomandibular joint pain and associated magnetic resonance findings: a retrospective study with a control group

Background To better understand and evaluate clinical usefulness of magnetic resonance imaging (MRI) in diagnosis and treatment of temporomandibular disorders (TMD), parameters for the evaluation are useful. Purpose To assess a clinically suitable staging system for evaluation of MRI of the temporomandibular joint (TMJ) and correlate the findings with age and some clinical symptoms of the TMJ. Material and Methods Retrospective analysis of 79 consecutive patients with clinical temporomandibular disorder or diagnosed inflammatory arthritis. Twenty-six healthy volunteers were included as controls. Existing data included TMJ pain, limited mouth opening (<30 mm) and corresponding MRI evaluations of the TMJs. Results The patients with clinical TMD complaints had statistically significantly more anterior disc displacement (ADD), disc deformation, caput flattening, surface destructions, osteophytes, and caput edema diagnosed by MRI compared to the controls. Among the arthritis patients, ADD, effusion, caput flattening, surface destructions, osteophytes, and caput edema were significantly more prevalent compared to the healthy volunteers. In the control group, disc deformation and presence of osteophytes significantly increased with age, and a borderline significance was found for ADD and surface destructions on the condylar head. No statistically significant associations were found between investigated clinical and MRI parameters. Conclusion This study presents a clinically suitable staging system for comparable MRI findings in the TMJs. Our results indicate that some findings are due to age-related degenerative changes rather than pathological changes. Results also show that clinical findings such as pain and limited mouth opening may not be related to changes diagnosed by MRI

Health-related quality of life and sleep quality after 12 months of treatment in non-severe obstructive sleep apnea: A randomized clinical trial with Continuous Positive Airway Pressure and Mandibular Advancement Splints

In this randomized controlled trial, patients with non-severe obstructive sleep apnea (OSA) were treated with continuous positive airway pressure (CPAP) or a twin block mandibular advancement splint (MAS). The primary objective was to compare how CPAP and MAS treatments change the health-related quality of life (HRQoL) and self-reported sleep quality of patients after 12 months of treatment. In total, 104 patients were recruited: 55 were allocated to the CPAP-treatment group and 49 to the MAS-treatment group. We used the SF36 questionnaire to evaluate HRQoL and the Pittsburgh Sleep Quality Index (PSQI) to evaluate sleep quality. All patients were included in the intention-to-treat analyses. These analyses showed improvements in the SF36 physical component score (from 48.8 ± 7.6 at baseline to 50.5 ± 8.0 at follow-up, p = 0.03) in the CPAP treatment group and in the mental component score (from 44.9 ± 12.1 to 49.3 ± 9.2, p = 0.009) in the MAS treatment group. The PSQI global score improved in both the CPAP (from 7.7 ± 3.5 to 6.6 ± 2.9, p = 0.006) and the MAS (8.0 ± 3.1 to 6.1 ± 2.6, p 0.05) in any analysis. The improvement in the SF36 vitality domain moderately correlated to the improvement in the PSQI global score in both groups (CPAP: |r| = 0.47, p < 0.001; MAS: |r| = 0.36, p = 0.01). In the MAS treatment group, we also found a weak correlation between improvements in the SF36 mental component score and PSQI global score (|r| = 0.28, p = 0.05). In conclusion, CPAP and MAS treatments lead to similar improvements in the HRQoL and self-reported sleep quality in non-severe OSA. Improvements in aspects of HRQoL seems to be moderately correlated to the self-reported sleep quality in both CPAP and MAS treatment

Friedman Score in Relation to Compliance and Treatment Response in Nonsevere Obstructive Sleep Apnea

Nonsevere obstructive sleep apnea (OSA) is most often treated with a continuous positive airway pressure (CPAP) device or a mandibular advancement splint (MAS). However, patient compliance with these treatments is difficult to predict. Improvement in apnea-hypopnea index (AHI) is also somewhat unpredictable in MAS treatment. In this study, we investigated the association between Friedman tongue position score (Friedman score) and both treatment compliance and AHI improvement in patients with nonsevere OSA receiving CPAP or MAS treatment. 104 patients with nonsevere OSA were randomly allocated to CPAP or MAS treatment and followed for 12 months. Data were collected through a medical examination, questionnaires, sleep recordings from ambulatory type 3 polygraphic sleep recording devices, and CPAP recordings. Associations between Friedman score, treatment compliance, and AHI improvement were analysed with logistic regression analyses. Friedman score was not associated with treatment compliance (odds ratio [OR]: 0.85, 95% confidence interval [CI]: 0.59–1.23), or AHI improvement (OR: 1.05, 95% CI: 0.62–1.76) in the overall study sample, the CPAP treatment group, or the MAS treatment group. Adjustment for socioeconomic factors, body mass index, and tonsil size did not significantly impact the results. Although Friedman score may predict OSA severity and contribute to the prediction of success in uvulopalatopharyngoplasty, we found no association between Friedman score and treatment compliance in patients with nonsevere OSA receiving CPAP or MAS treatment, nor did we find any association between Friedman score and AHI improvement. Factors other than Friedman score should be considered when deciding whether a patient with nonsevere OSA should be treated with CPAP or MAS