Rationale for a Combination Therapy with the STAT5 Inhibitor AC-4-130 and the MCL1 Inhibitor S63845 in the Treatment of FLT3-Mutated or TET2-Mutated Acute Myeloid Leukemia.

The FMS-like tyrosine kinase 3 (FLT3) gene is mutated in one-third of patients with de novo acute myeloid leukemia (AML). Mutated FLT3 variants are constitutively active kinases signaling via AKT kinase, MAP kinases, and STAT5. FLT3 inhibitors have been approved for the treatment of FLT3-mutated AML. However, treatment response to FLT3 inhibitors may be short-lived, and resistance may emerge. Compounds targeting STAT5 may enhance and prolong effects of FLT3 inhibitors in this subset of patients with FLT3-mutated AML. Here STAT5-inhibitor AC-4-130, FLT3 inhibitor midostaurin (PKC412), BMI-1 inhibitor PTC596, MEK-inhibitor trametinib, MCL1-inhibitor S63845, and BCL-2 inhibitor venetoclax were assessed as single agents and in combination for their ability to induce apoptosis and cell death in leukemic cells grown in the absence or presence of bone marrow stroma. Synergistic effects on cell viability were detected in both FLT3-mutated and FLT3-wild-type AML cells treated with AC-4-130 in combination with the MCL1 inhibitor S63845. AML patient samples with a strong response to AC-4-130 and S63845 combination treatment were characterized by mutated FLT3 or mutated TET2 genes. Susceptibility of AML cells to AC-4-130, PTC596, trametinib, PKC412, and venetoclax was altered in the presence of HS-5 stroma. Only the MCL1 inhibitor S63845 induced cell death with equal efficacy in the absence or presence of bone marrow stroma. The combination of the STAT5-inhibitor AC-4-130 and the MCL1 inhibitor S63845 may be an effective treatment targeting FLT3-mutated or TET2-mutated AML.

Volumetric analysis of chin and mandibular retromolar region as donor sites for cortico-cancellous bone blocks.

AIM: To test whether the mandibular retromolar region renders different results from the chin region with respect to the amount of bone available for the harvesting of block grafts. MATERIAL AND METHODS: Sixty cone beam computed tomography (CBCT) scans of mandibles of adult patients without pathologic findings in the chin and retromolar region were included. According to the number of mandibular teeth, 20 CBCT data sets were allocated to each of the following groups: group M1: dentition 36-46; group M2: dentition 37-47; and group M3: dentition 38-48. For the potential donor sites in the chin and the retromolar regions, the volume (VChin , VRetro ), the length (LChin , LRetro ), the height (HChin , HRetro ) and the width (HChin , HRetro ) were assessed using a computer software. Moreover, the chin was examined for the presence and the localization of the mandibular incisive canal. To compare the donor sites in the chin and in the retromolar regions, the quotients VRetro /VChin , LRetro /LChin , HRetro /HChin and WRetro /WChin were calculated and tested using the Wilcoxon signed-rank test or the sign test. RESULTS: The mean bone volume VChin measured 3.5 ± 1.3 cm(3) (SD), whereas the overall VRetro amounted to 1.8 ± 1.1 cm(3) (SD). VRetro amounted to 2.6 ± 1.4 cm(3) (SD) in the group M1, 1.8 ± 0.5 cm(3) in the group M2 and 1.0 ± 0.4 cm(3) in the group M3. For the group M1, VRetro /VChin measured 82 ± 39% (P = 0.036). VRetro /VChin reached 57 ± 20% in the group M2 and 32 ± 12% in the group M3 (P < 0.001). The mandibular incisive canal was detected in 97% of the CBCT scans. The distance between the mandibular incisive canal and the apices of the central incisors measured 10.5 ± 3.5 mm. CONCLUSION: The amount of bone available for the harvesting of cortico-cancellous blocks in the chin region was superior in comparison with the mandibular retromolar region. In the absence of the second and the third molars, the amount of bone harvestable in the retromolar region reached approximately 80% of the bone volume available in the chin region. In the majority of the cases, the mandibular incisive canal was detected within the donor site in the chin region.

Comparative release of growth factors from PRP, PRF, and advanced-PRF.

OBJECTIVES: The use of platelet concentrates has gained increasing awareness in recent years for regenerative procedures in modern dentistry. The aim of the present study was to compare growth factor release over time from platelet-rich plasma (PRP), platelet-rich fibrin (PRF), and a modernized protocol for PRF, advanced-PRF (A-PRF). MATERIALS AND METHODS: Eighteen blood samples were collected from six donors (3 samples each for PRP, PRF, and A-PRF). Following preparation, samples were incubated in a plate shaker and assessed for growth factor release at 15 min, 60 min, 8 h, 1 day, 3 days, and 10 days. Thereafter, growth factor release of PDGF-AA, PDGF-AB, PDGF-BB, TGFB1, VEGF, EGF, and IGF was quantified using ELISA. RESULTS: The highest reported growth factor released from platelet concentrates was PDGF-AA followed by PDGF-BB, TGFB1, VEGF, and PDGF-AB. In general, following 15-60 min incubation, PRP released significantly higher growth factors when compared to PRF and A-PRF. At later time points up to 10 days, it was routinely found that A-PRF released the highest total growth factors. Furthermore, A-PRF released significantly higher total protein accumulated over a 10-day period when compared to PRP or PRF. CONCLUSION: The results from the present study indicate that the various platelet concentrates have quite different release kinetics. The advantage of PRP is the release of significantly higher proteins at earlier time points whereas PRF displayed a continual and steady release of growth factors over a 10-day period. Furthermore, in general, it was observed that the new formulation of PRF (A-PRF) released significantly higher total quantities of growth factors when compared to traditional PRF. CLINICAL RELEVANCE: Based on these findings, PRP can be recommended for fast delivery of growth factors whereas A-PRF is better-suited for long-term release.

Effect of Titanium and Zirconium Oxide Microparticles on Pro-Inflammatory Response in Human Macrophages under Induced Sterile Inflammation: An In Vitro Study.

The wear-debris particles released by shearing forces during dental implant insertion may contribute to inflammatory reactions or osteolysis associated with peri-implantitis by stimulating inflammasome-activation. The study aim was to examine cytotoxic and pro-inflammatory effects of titanium (TiO2) and zirconia (ZrO2) particles in macrophages regarding their nature/particle concentration over time under sterile lipopolysaccharide (LPS) inflammation. Macrophages were exposed to TiO2 and ZrO2 particles (≤5 µm) in cell culture. Dental glass was used as inert control and LPS (1 µg/mL) was used to promote sterile inflammation. Cytotoxicity was determined using MTT assays and cytokine expression of TNF-α, IL-1ß and IL-6 was evaluated by qRT-PCR. Data were analyzed using Student's t-test and ANOVA (p ≤ 0.05). Cytotoxicity was significantly increased when exposed to higher concentrations of glass, TiO2 and ZrO2 (≥107 particles/mL) compared to controls (p ≤ 0.05). Macrophages challenged with TiO2 particles expressed up to ≈3.5-fold higher upregulation than ZrO2 from 12 to 48 h. However, when exposed to LPS, TiO2 and ZrO2 particle-induced pro-inflammatory gene expression was further enhanced (p ≤ 0.05). Our data suggest that ZrO2 particles produce less toxicity/inflammatory cytokine production than TiO2. The present study shows that the biological reactivity of TiO2 and ZrO2 depends on the type and concentration of particles in a time-dependent manner.