Severe Compromise of Preosteoblasts in a Surgical Mouse Model of Bisphosphonate-Associated Osteonecrosis of the Jaw.

Objectives: The effect of amino-bisphosphonates on osteoblastic lineage and its potential contribution to the pathogenesis of bisphosphonate-associated osteonecrosis of the jaw (BONJ) remain controversial. We assessed the effects of zoledronic acid (ZOL) on bone and vascular cells of the alveolar socket using a mouse model of BONJ. Material and Methods: Thirty-two mice were treated twice a week with either 100 μg/kg of ZOL or saline for 12 weeks. The first left maxillary molar was extracted at the third week. Alveolar sockets were assessed at both 3 weeks (intermediate) and 9 weeks (long-term) after molar extraction by semi-quantitative histomorphometry for empty lacunae, preosteoblasts (Osterix), osteoclasts (TRAP), and pericyte-like cells (CD146). Also, the bone microarchitecture was assessed by micro-CT. Results: Osteonecrotic-like lesions were observed in 21% of mice. Moreover, a decreased number of preosteoblasts contrasted with the increased number of osteoclasts at both time points. In addition, osteoclasts display multinucleation and detachment from the endosteal surface. Furthermore, the number of pericyte-like cells increased at the intermediate time point. The alveolar bone mass increased exclusively with long-term ZOL treatment. Conclusion: The severe imbalance between bone-forming cells and bone-resorbing cells showed in this study could contribute to the pathogenesis of BONJ

Prospective purification of perivascular presumptive mesenchymal stem cells from human adipose tissue:process optimization and cell population metrics across a large cohort of diverse demographics

BACKGROUND: Adipose tissue is an attractive source of mesenchymal stem cells (MSC) as it is largely dispensable and readily accessible through minimally invasive procedures such as liposuction. Until recently MSC could only be isolated in a process involving ex-vivo culture and their in-vivo identity, location and frequency remained elusive. We have documented that pericytes (CD45-, CD146+, and CD34-) and adventitial cells (CD45-, CD146-, CD34+) (collectively termed perivascular stem cells or PSC) represent native ancestors of the MSC, and can be prospectively purified using fluorescence activated cell sorting (FACS). In this study we describe an optimized protocol that aims to deliver pure, viable and consistent yields of PSC from adipose tissue. We analysed the frequency of PSC within adipose tissue, and the effect of patient and procedure based variables on this yield. METHODS: Within this twin centre study we analysed the adipose tissue of n = 131 donors using flow cytometry to determine the frequency of PSC and correlate this with demographic and processing data such as age, sex, BMI and cold storage time of the tissue. RESULTS: The mean number of stromal vascular fraction (SVF) cells from 100 ml of lipoaspirate was 34.4 million. Within the SVF, mean cell viability was 83 %, with 31.6 % of cells being haematopoietic (CD45+). Adventitial cells and pericytes represented 33.0 % and 8 % of SVF cells respectively. Therefore, a 200 ml lipoaspirate would theoretically yield 23.2 million viable prospectively purified PSC - sufficient for many reconstructive and regenerative applications. Minimal changes were observed in respect to age, sex and BMI suggesting universal potential application. CONCLUSIONS: Adipose tissue contains two anatomically and phenotypically discreet populations of MSC precursors – adventitial cells and pericytes – together referred to as perivascular stem cells (PSC). More than 9 million PSC per 100 ml of lipoaspirate can be rapidly purified to homogeneity using flow cytometry in clinically relevant numbers potentially circumventing the need for purification and expansion by culture prior to clinical use. The number and viability of PSC are minimally affected by patient age, sex, BMI or the storage time of the tissue, but the quality and consistency of yield can be significantly influenced by procedure based variables. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13287-016-0302-7) contains supplementary material, which is available to authorized users

Testis sparing surgery for Leydig cell tumors: New three cases and review of the current literature

Introduction: Leydig cells tumors of the testis are uncommon, representing between 1 and 3% of the testicular tumors and for which the natural history and therapy are debated between radical orchitectomy and organ-sparing surgery. Subjects and methods: We report three new cases of Leydig cells tumors, treated in our department and we discuss the clinical, diagnostic and therapeutic aspects of this uncommon tumor. Results: The mean patient age was 29 years (23–37 years). Medical referral was for a testicular pain in two and gynecomastia in one case. All patients were treated surgically, through an inguinal incision and the procedure included clamping of the spermatic cord. During surgery, frozen section were analyzed in two cases and the tumor enucleation with organ-sparing surgery was performed. The other patient had an unilateral orchidectomy and controlateral tumorectomy. The mean follow-up was 40 months with no local recurrence. Conclusion: Patients diagnosed with Leydig cells tumors have a good prognosis; this study shows the safety of conservative surgery treatment, provided it is subsequently followed by close surveillance, as it preserves maximum fertility, and these tumors usually have a favorable prognosis

Chemical composition, cooking quality, texture and consumer acceptance of pasta with Eruca vesicaria leaves

peer reviewedIn this study, pasta with dried Eruca vesicaria leaves commonly known as Rocket salad and spinach leaves flours were prepared. Samples were evaluated for its chemical composition, cooking quality, textural, colour and consumer acceptance. Results showed that pasta with Eruca vesicaria had the highest dietary fibre content (5.30–9.50 g/100 g) and the lowest fat content (2.13–2.80 g/100 g). The optimally cooked pasta with 5% of leaves (stored 30 days) and pasta with 10% of leaves (stored 14 days) have good cooking quality with cooking loss ≤8%. The green colour and textural characteristics of pastas were stable during all period at 4 °C. Textural characteristics of pasta with rocket and spinach were similar. Pasta with 10% of rocket leaves has presented the highest overall acceptability score