Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability

Abstract

Objectives

The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs).

Materials and methods

Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1  % fetal bovine serum (low-serum) or α-MEM containing 10  % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation.

Results

Low-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation.

Conclusions

MBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium.

Clinical relevance

These findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.

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Biomarkers in diagnosis and therapy of oral squamous cell carcinoma: A review of the literature

Publication date: Available online 16 February 2017
Source:Journal of Cranio-Maxillofacial Surgery
Author(s): Sebastian Blatt, Maximilian Krüger, Thomas Ziebart, Keyvan Sagheb, Eik Schiegnitz, Elisabeth Goetze, Bilal Al-Nawas, Andreas Max Pabst
Oral squamous cell carcinoma (OSCC) represents the sixth most common cancer, accounting for 2-4% of all malignancies worldwide. The overall survival rate of less than 60% remains generally poor, with prognosis heavily relying on the TNM staging system. Tumor size as well as the presence and extent of lymph node metastases are widely recognized as the most important predictors. However, the underlying mechanisms that lead to an aggressive phenotype are not yet fully understood. Therefore, possible biomarkers are much in need to predict prognosis, to help individualize therapy approaches, and to overcome possible resistance mechanisms.Despite a multitude of recently published biomarkers for OSCC, there is still an ongoing debate regarding their implementation in the clinical workflow. Thus, a systematic literature search via PubMed was performed to update the current literature with the latest evidence. In total, 128 studies were included and over 100 different biomarkers evaluated with reference to their influence of survival, tumor recurrence, advanced grading and lymph node metastasis.In this review, we highlight the important molecular mechanism underlying possible markers in tissue, blood or saliva samples for OSCC. As a major result, no clinical trials could be obtained to prove clinical importance of the validated predictors for survival, tumor recurrence, lymph node metastasis and therapy resistance. Therefore, further clinical investigations are much needed.



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Anti-Gal and anti-Neu5Gc responses in nonimmunosuppressed patients following treatment with rabbit anti-thymocyte polyclonal IgGs.

Background: Polyclonal anti-human thymocyte rabbit IgGs (anti-thymocyte globulin, ATG) are popular immunosuppressive drugs used to prevent or treat organ or bone-marrow allograft rejection, graft versus host disease, and autoimmune diseases. However, animal-derived glycoproteins are also strongly immunogenic and rabbit ATG induces serum sickness disease in almost all patients without additional immunosuppressive drugs, as seen in the START trial of ATG therapy in new-onset type 1 diabetes. Methods: Using ELISA, we analyzed serial sera from the START study to decipher the various anti-ATG specificities developed by the patients in this study: anti-total ATG, but also anti-galactose-[alpha]1-3-galactose (Gal) and anti-Neu5Gc antibodies, 2 xeno-carbohydrate epitopes present on rabbit IgG glycans and lacking in humans. Results: We show that diabetic patients have substantial levels of preexisting antibodies of the 3 specificities, before infusion, but of similar levels as healthy individuals. ATG treatment resulted in highly significant increases of both IgM (for anti-ATG and anti-Neu5Gc) and IgG (for anti-ATG, -Gal, and -Neu5Gc), peaking at 1 month and still detectable 1 year postinfusion. Conclusions: Treatment with rabbit polyclonal IgGs in the absence of additional immunosuppression results in a vigorous response against Gal and Neu5Gc epitopes, contributing to an inflammatory environment that may compromise the efficacy of ATG therapy. The results also suggest using IgGs lacking these major xeno-antigens may improve safety and efficacy of ATG treatment. Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved.

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Long-term follow up of humoral immune status in adult lung transplant recipients.

Background: Lung transplant recipients have an increased risk for infections in the posttransplant period due to immunosuppressive therapy. Protection against infections can be achieved through vaccination, but the optimal vaccination schedule in lung transplant recipients is unknown. Data on long-term immunological follow up and vaccination responses after lung transplantation are scarce. Methods: Here we present long-term immunological follow up of a cohort of 55 lung transplant recipients. This includes detailed antibody responses after 23-valent pneumococcal polysaccharide vaccination (23vPPV). Results: All patients were vaccinated with 23vPPV prior to transplantation. Median follow-up after transplantation was 6.6 years (379 patient-years). Following transplantation, there is a significant decrease of all immunoglobulins, IgG subclasses and pneumococcal polysaccharide antibodies. After the first year posttransplantation there is a gradual increase of all immunoglobulins and IgG subclasses, but values were always significantly lower than in the pretransplant period. After a median of 4.4 years posttransplantation patients were revaccinated with 23vPPV. The pneumococcal polysaccharide antibody response was impaired in 87% of patients (ie, antibody titer above cutoff and 2-fold increase between pre and postvaccination values for

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Imaging Leukocyte Responses in the Kidney.

The kidney can be negatively affected by a range of innate and adaptive immune responses, resulting in alterations in the functions of the kidney and, in some cases, progression to renal failure. In many of these responses, infiltration of blood-borne leukocytes into the kidney is central to the response. In addition, a large population of mononuclear phagocytes resident in the kidney can modulate these responses. A great deal of research has investigated both the mechanisms of leukocyte recruitment to the kidney and the actions of immune cells resident within the kidney. Due to the dynamic nature of the processes whereby leukocytes enter sites of inflammation, in vivo imaging has been 1 of the key approaches used for understanding leukocyte recruitment as it occurs throughout the body, and this is also true of the kidney. However, imaging this organ and its complicated microvasculature during different forms of renal pathology presents a unique set of challenges. In this review, we examine the approaches used for intravital imaging of the kidney, and summarise the insights gained from these studies regarding the mechanisms of leukocyte entry into kidney during inflammation, and the actions of immune cells within this organ. Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved.

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The differential effect of apyrase treatment and hCD39 overexpression on chronic renal fibrosis following ischemia reperfusion injury.

Background: Renal ischemia reperfusion injury (IRI) leads to acute kidney injury and renal fibrosis. CD39 is a key purinergic enzyme in the hydrolysis of ATP and increased CD39 enzymatic activity protects from acute IRI but its effect on renal fibrosis is not known. Methods: Using a mouse model of unilateral renal IRI, the effects of increased CD39 activity (using soluble apyrase and mice expressing human CD39 transgene) on acute and chronic renal outcomes were examined. Nucleotide (ATP, ADP, AMP) and nucleoside (adenosine and inosine) levels were quantified by HPLC. Soluble apyrase reduced acute renal injury at 24 hours and renal fibrosis at 4 weeks post-IRI, compared to vehicle-treated mice. Results: Soluble apyrase reduced renal ATP, ADP and AMP, but not adenosine levels, during ischemia. In comparison with wild-type littermates, hCD39 transgenic mice were protected from acute renal injury at 24 hours, but had increased renal fibrosis at 4 weeks post-IRI. hCD39 transgene expression was localized to the vascular endothelium at baseline, and did not affect total renal nucleotide and nucleoside levels during ischemia. However, hCD39 transgene was more widespread at 4 weeks post-IRI and was associated with higher renal adenosine levels at 4 weeks post-IRI compared to wild-type littermates. Conclusion: A single dose of apyrase administration before IRI protects from both acute and chronic renal injury and may have clinical application in protection from ischemic-induced renal injury. Furthermore, transgenic expression of hCD39 is associated with increased renal fibrosis after ischemia. Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved.

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Japanese guidelines for allergic rhinitis 2017

Publication date: Available online 15 February 2017
Source:Allergology International
Author(s): Kimihiro Okubo, Yuichi Kurono, Keiichi Ichimura, Tadao Enomoto, Yoshitaka Okamoto, Hideyuki Kawauchi, Harumi Suzaki, Shigeharu Fujieda, Keisuke Masuyama
Like asthma and atopic dermatitis, allergic rhinitis is an allergic disease, but of the three, it is the only type I allergic disease. Allergic rhinitis includes pollinosis, which is intractable and reduces quality of life (QOL) when it becomes severe. A guideline is needed to understand allergic rhinitis and to use this knowledge to develop a treatment plan. In Japan, the first guideline was prepared after a symposium held by the Japanese Society of Allergology in 1993. The current 8th edition was published in 2016, and is widely used today.To incorporate evidence based medicine (EBM) introduced from abroad, the most recent collection of evidence/literature was supplemented to the Practical Guideline for the Management of Allergic Rhinitis in Japan 2016. The revised guideline includes assessment of diagnosis/treatment and prescriptions for children and pregnant women, for broad clinical applications. An evidence-based step-by-step strategy for treatment is also described. In addition, the QOL concept and cost benefit analyses are also addressed. Along with Allergic Rhinitis and its Impact of Asthma (ARIA), this guideline is widely used for various clinical purposes, such as measures for patients with sinusitis, childhood allergic rhinitis, oral allergy syndrome, and anaphylaxis and for pregnant women. A Q&A section regarding allergic rhinitis in Japan was added to the end of this guideline.



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