Role of Adult Tissue-Derived Pluripotent Stem Cells in Bone Regeneration.

BACKGROUND: Bone marrow-derived mononuclear cells (BM-MNC) consist of a heterogeneous mix of mesenchymal stem cells (MSC), hematopoietic progenitor cells (HPC), endothelial progenitor cells (EPC), monocytes, lymphocytes and pluripotent stem cells. Whereas the importance of MSC and EPC has been well documented in bone healing and regeneration studies, the role of pluripotent stem cells is still poorly understood. In the present study we evaluated if and how Very Small Embryonic Like cells (VSEL), isolated from rat BM-MNC, contribute to bone healing. METHODS: Large bone defects were made in the femurs of 38 Sprague Dawley female rats and treated with ß-TCP scaffold granules seeded with male VSEL; BM-MNC, VSEL-depleted BM-MNC or scaffold alone, and bone healing was evaluated at 8 weeks post-surgery. RESULTS: Bone healing was significantly increased in defects treated with VSEL and BM-MNC, compared to defects treated with VSEL-depleted BM-MNC. Donor cells were detected in new bone tissue, in all the defects treated with cells, and in fibrous tissue only in defects treated with VSEL-depleted BM-MNC. The number of CD68+ cells was the highest in the VSEL-depleted group, whereas the number of TRAP positive cells was the lowest in this group. CONCLUSIONS: Based on the results, we can conclude that VSEL play a role in BM-MNC induced bone formation. In our rat femur defect model, in defects treated with VSEL-depleted BM-MNC, osteoclastogenesis and bone formation were decreased, and foreign body reaction was increased.

Electrical stimulation shifts healing/scarring towards regeneration in a rat limb amputation model.

Different species respond differently to severe injury, such as limb loss. In species that regenerate, limb loss is met with complete restoration of the limbs' form and function, whereas in mammals the amputated limb's stump heals and scars. In in vitro studies, electrical stimulation (EStim) has been shown to promote cell migration, and osteo- and chondrogenesis. In in vivo studies, after limb amputation, EStim causes significant new bone, cartilage and vessel growth. Here, in a rat model, the stumps of amputated rat limbs were exposed to EStim, and we measured extracellular matrix (ECM) deposition, macrophage distribution, cell proliferation and gene expression changes at early (3 and 7 days) and later stages (28 days). We found that EStim caused differences in ECM deposition, with less condensed collagen fibrils, and modified macrophage response by changing M1 to M2 macrophage ratio. The number of proliferating cells was increased in EStim treated stumps 7 days after amputation, and transcriptome data strongly supported our histological findings, with activated gene pathways known to play key roles in embryonic development and regeneration. In conclusion, our findings support the hypothesis that EStim shifts injury response from healing/scarring towards regeneration. A better understanding of if and how EStim controls these changes, could lead to strategies that replace scarring with regeneration.

Using the Cancer Risk Management Model to evaluate the health and economic impacts of cytology compared with human papillomavirus DNA testing for primary cervical cancer screening in Canada.

BACKGROUND: In Canada, discussion about changing from cytology to human papillomavirus (hpv) dna testing for primary screening in cervical cancer is ongoing. However, the Canadian Task Force on Preventive Health Care has not yet made a recommendation, concluding that the evidence is insufficient. METHODS: We used the cervical cancer and hpv transmission models of the Cancer Risk Management Model to study the health and economic outcomes of primary cytology compared with hpv dna testing in 14 screening scenarios with varying screening modalities and intervals. Projected cervical cancer cases, deaths, colposcopies, screens, costs, and incremental cost-effectiveness were evaluated. We performed sensitivity analyses for hpv dna test costs. RESULTS: Compared with triennial cytology from age 25, 5-yearly hpv dna screening alone from age 30 resulted in equivalent incident cases and deaths, but 55% (82,000) fewer colposcopies and 43% (1,195,000) fewer screens. At hpv dna screening intervals of 3 years, whether alone or in an age-based sequence with cytology, screening costs are greater, but at intervals of more than 5 years, they are lower. Scenarios on the cost-effectiveness frontier were hpv dna testing alone every 10, 7.5, 5, or 3 years, and triennial cytology starting at age 21 or 25 when combined with hpv dna testing every 3 years. CONCLUSIONS: Changing from cytology to hpv dna testing as the primary screening test for cervical cancer would be an acceptable strategy in Canada with respect to incidence, mortality, screening and diagnostic test volumes.

Gingival crevicular blood for screening of blood glucose level in patients with & without diabetes: a chair-side test.

OBJECTIVES: Diabetes is a pandemic disease with increasing prevalence and serious complications. Periodontitis being one of its presentation and is its sixth recognized complication. This study compares blood glucose levels in gingival crevicular blood of patients with and without diabetes elicited during routine periodontal probing and venous blood sample. METHODS: Seventy patients with moderate gingivitis and periodontitis positive for bleeding on probing were chosen. All the subjects were divided in two groups, group I consisted of 35 diabetic and group II of 35 non-diabetic subjects. Blood from the gingiva of the most inflamed site was collected with the test strip of a glucose self-monitoring device, and the blood glucose levels were measured. At the same time, intravenous blood was collected for measurement in a laboratory glucose analyzer. Gingival index and probing pocket depth were evaluated for each subject at same time. RESULTS: The mean GCB levels and VB derived from all samples were 156.07 ± 49.23 mg dl(-1) and 156 ± 49.89 mg dl(-1) , respectively, for diabetic group and 90.80 ± 11.07 and 93.41 ± 9.30 for non-diabetic group. In both the groups, the difference between GCB and VB glucose levels was non-significant (P > 0.005). Highly significant correlation between GCB and VB (r = 0.972 for diabetic and r = 0.721 for non-diabetic) in both the groups was found. CONCLUSION: The data from this study show that GCB collected during diagnostic periodontal examination can be an excellent source for estimation of blood sugar or glucometric analysis. This technique is also suitable for routine screening of diabetic and early diagnosis of unknown diabetic cases.

A randomized two way cross over study for comparison of absorption of vitamin D3 buccal spray and soft gelatin capsule formulation in healthy subjects and in patients with intestinal malabsorption.

BACKGROUND: Vitamin D deficiency has been proposed to contribute to the development of malabsorption diseases. Despite this, the vitamin D status of these patients is often neglected. The objective of the present work was to compare the absorption of vitamin D3 through the oral route by comparing a 1000 IU soft gelatin capsule and a 500 IU buccal spray (delivering 1000 IU in two spray shots) in healthy subjects and in patients with malabsorption disease. METHODS: An open label, randomized, two-periods, two-way cross over study was conducted, first in healthy subjects (n = 20) and then in patients with malabsorption syndrome (n = 20). The study participants were equally divided and received either of the treatments (buccal spray, n = 7; soft gelatin capsule, n = 7; control, n = 6) in Period I for 30 days. After washout of another 30 days, the treatments were changed in crossover fashion in Period II. Fasting blood samples were collected to measure baseline 25-hydroxyvitamin D [25(OH)D] levels in all participants at day 0 (Screening visit), day 30 (completion of period I), day 60 (end of wash out and initiation of period II) and day 90 (completion of period II). Safety was evaluated by hematology and biochemistry analyses. Statistical analyses was performed using differences of mean and percentage change from baseline of 25(OH)D levels between two formulation by two tailed Paired t-test with 95% confidence interval. RESULTS: In healthy subjects, the mean increase in serum 25(OH)D concentration was 4.06 (95% CI 3.41, 4.71) ng/ml in soft gelatin capsule group and 8.0 (95% CI 6.86, 9.13) ng/ml in buccal spray group after 30 days treatment (p < 0.0001). In patients with malabsorption disease, the mean increase in serum 25(OH)D concentration was 3.96 (95% CI 2.37, 5.56) ng/ml in soft gelatin capsule group and 10.46 (95% CI 6.89, 14.03) ng/ml in buccal spray group (p < 0.0001). CONCLUSION: It can be concluded from the results that the buccal spray produced a significantly higher mean serum 25(OH)D concentration as compared to the soft gelatin capsule, in both healthy subjects as well as in patients with malabsorption syndrome over a period of 30 days administration in a two way cross over study. Treatments were well tolerated by both subject groups TRIAL REGISTRATION: CTRI/2013/06/003770.

Oral IL-10 gene delivery in a microsphere-based formulation for local transfection and therapeutic efficacy in inflammatory bowel disease.

The objective of this study was to examine the potential of oral interleukin-10 (IL-10) gene therapy for the treatment of inflammatory bowel disease (IBD). Nanoparticles-in-microsphere oral system (NiMOS) was formulated with murine IL-10-expressing plasmid DNA in type-B gelatin nanoparticles, which were further encapsulated in poly(epsilon-caprolactone) microsphere matrix. Upon oral administration in an acute colitis model, IL-10 expression in the large intestine was measured by quantitative real-time PCR and ELISA. The locally expressed IL-10 was able to suppress the levels of proinflammatory cytokines, such as IFN-gamma, TNF-alpha, IL-1alpha, IL-1beta and IL-12, as well as certain chemokines. The therapeutic benefits of transfected IL-10 were further demonstrated by an increase in body weight, favorable clinical activity score, restoration in colon length and weight, and suppression of inflammatory response as assessed by tissue histological analysis and myeloperoxidase activity. The results of this study provide highly encouraging evidence of oral gene delivery and transfection and potential utility in IBD therapy.

Project Dil: a co-ordinated Primary Care and Community Health Promotion Programme for reducing risk factors of coronary heart disease amongst the South Asian community of Leicester--experiences and evaluation of the project.

OBJECTIVES: To improve the effectiveness of primary and secondary prevention of coronary heart disease (CHD) in volunteer Leicestershire general practices with a high percentage of South Asian patients. To increase the awareness of lifestyle risk factors amongst the South Asian community with means of reducing CHD. DESIGN: Development and implementation of: A CHD training and awareness programme for health care professionals. Organizational change to ensure adoption of an effective secondary prevention programme for general practice. A public awareness campaign including a peer education programme for the South Asian community of Leicestershire. Interim evaluation using participation data, user satisfaction and organizational development of primary care is reported. RESULTS: A multi-disciplinary training programme accessed by 88% of staff from 23 volunteer practices has been developed and implemented. Organizational development in practices including CHD registers, action planning and establishing CHD clinics has taken place at a much faster pace than comparable non-Project Dil practices. An externally accredited peer education programme has been completed by 45 community volunteers. Within 6 months of completion, a total of 54 peer education sessions accessed by over 2,000 people from the Asian community had taken place. Project Dil, although initially funded only for 2 years, has now been adopted by Leicestershire Health Services (via a Primary Care Trust) as a mainstream programme. CONCLUSION: Project Dil has made considerable progress in engaging and driving changes in general practices to facilitate improvement in CHD management. In parallel, the project has developed a community education programme, including the use of peer education. The project is now hosted by a Primary Care Trust on behalf of Leicestershire Health Services. External evaluation of the peer education programme has commenced.