Search Results (21)

Mesenchymal stromal cells of the tumor microenvironment (TME) play a significant role in the progression of multiple myeloma (MM). The cells of the TME demonstrate resistance to treatment, thereby creating a favorable environment for disease relapse. The status of the TME during remission is poorly understood. An association between treatment response and TME status (including signaling pathways) has been suggested. One of the key players in the establishment of the MM TME is WNT signaling. In this study, we evaluated the expression of WNT family proteins in the TME and MM cells to assess their potential as TME markers and predictors of treatment response. A bioinformatic analysis of normal and malignant plasma cells, combined with an analysis of published data, revealed the following differentially expressed WNT genes: WNT5A, WNT10B, CTNNB1, and WNT3A. Immunohistochemical staining with the antibodies against the proteins encoded by the genes was conducted on trephine biopsy samples of bone marrow from healthy donors and patients with different responses to therapy. A quantitative analysis of the immunohistochemical data revealed differences in the amounts of WNT3A, WNT5A, WNT10B, and β-catenin proteins in the bone marrow before treatment depending on the subsequent responses of the patients to therapy. Multiplex fluorescent immunohistochemical staining with tyramide signal amplification revealed that WNT3A was predominantly present in mesenchymal stromal cells, whereas WNT5A and WNT10B were primarily observed in plasma cells. β-catenin was detected in both cell types. We analyzed the mRNA levels of the WNT gene family and CTNNB1 in MSC cultures from healthy donors and patients using qPCR. These genes were differentially expressed in MSC cultures derived from patients and healthy donors, as well as between patients grouped according to their response to therapy. Therefore, WNT proteins and β-catenin can be considered potential markers to assess the state of the tumor niche. Full article

Myelodysplastic neoplasms (MDS) represent a heterogeneous group of neoplastic bone marrow disorders. A crucial component in regulating bone marrow cell apoptosis is the B-cell CLL/lymphoma 2 (BCL-2) protein. This retrospective study aimed to assess BCL-2 expression by immunohistochemistry in trephine biopsy specimens from 76 patients diagnosed with MDS. The obtained retrospective results were correlated with clinical parameters, including age, sex, MDS subtype, IPSS, IPSS-R, bone marrow blast percentage, Ogata score, response to treatment, blood morphology parameters, and overall survival (OS). The median follow-up duration was 16 months. During the observation period, 58 patients died (median OS of this group: 14.6 months), and 25 patients experienced progression to acute myeloid leukemia. The median BCL-2 expression assessed using the Histoscore (H-score) was 10. Patients with BCL-2 expression below 10 had better survival outcomes than those with expression ≥ 10. Furthermore, patients without detectable BCL-2 expression had significantly better survival compared to those with detectable BCL-2 expression (p = 0.0084). Higher BCL-2 expression was significantly associated with high and very high cytogenetic risk, as defined by IPSS-R. BCL-2 immunohistochemistry should be viewed as a complementary biomarker that, when integrated with IPSS-R and mutational data, could refine therapeutic algorithms. Full article

Background/Objectives: There are limited data regarding immunohistochemical profiling of immune cells in bone marrow trephine biopsies of patients with high-risk myelodysplastic syndromes (HR-MDS). Methods: We sought to objectively quantify, with the use of digital pathology, the density (cells/mm2) of the prominent adaptive immunity cell populations in sixty-four (64) bone marrow trephine biopsies of HR-MDS patients receiving 5-Azacytidine. We focused on CD3(+) T cells, CD8(+) cytotoxic T cells (Tc), helper T cells (Th), Foxp3(+) regulatory T cells (Tregs), CD20(+) B-cells and CD138(+) plasma cells and evaluated the presence and the number of lymphoid aggregates. A control group of twenty “non-MDS” patients was included in the study. Results: We identified a significant decrease in adaptive immune cell densities in the HR-MDS patients compared to the non-MDS controls. Increased T and Th cell densities correlated with the response to 5-Azacytidine (5-AZA) treatment. Higher T, Tc, Th and plasma cells densities and low B, Tregs and Tregs/T cells ratios correlated with increased overall survival. Reduced Tregs, Tregs/T cells, Tregs/Tc and plasma cells showed improved leukemia-free survival. A modified IPSS-R (IPSS-R-I), combining the initial IPSS-R with the immune populations’ parameters, improved overall survival and showed a double-fold increase in Cox calculated hazard ratios. Conclusions: Immunohistochemical bone marrow immune profiling represents a powerful and easily useable tool for investigating the possible role of bone marrow immune microenvironment in the pathogenesis and progression of MDS, but also its association with the response to 5-AZA treatment and clinical outcomes. Full article

Background/Objectives: Despite the increased use of new resorbable magnesium membranes, there are no reported cases or studies on the use of resorbable magnesium membranes in combination with bone grafts for alveolar ridge preservation (ARP) in cases with severe buccal bone wall dehiscence. This case report aimed to evaluate the effectiveness of the magnesium membrane shield technique in conjunction with bone grafting for ARP, assessing both clinical outcomes and histological bone regeneration. Methods: A 44-year-old female patient presented with a vertical fracture on tooth 24 (FDI Notation System) accompanied with complete destruction of the buccal bone wall. The treatment plan included tooth extraction, ARP using a combination of anorganic bovine bone and autologous bone grafting, and the application of a magnesium membrane as a shield to the pre-existing buccal wall. Six months post-procedure, a bone biopsy was taken from the implant site using a trephine bur. Results: Clinical and radiological evaluations six months after the procedure demonstrated sufficient bone volume for implant placement. Additionally, in the next three months, soft tissue conditioning with a provisional crown resulted in an aesthetically and functionally satisfactory outcome. Histological analysis of the bone biopsy revealed well-formed new bone in direct contact with residual biomaterial, with no signs of inflammation. Osteocytes were clearly visible within the newly formed bone matrix, indicating successful bone maturation. Active osteoblasts were observed along the bone-biomaterial interface, suggesting ongoing bone remodeling and integration. Additionally, histomorphometric evaluation revealed 47% newly formed bone, 32% soft tissue, and 19% residual biomaterial. Conclusions: This case demonstrates the potential of the magnesium shield technique as an ARP technique in cases with severe buccal wall dehiscence. The technique yielded satisfactory clinical outcomes and promoted successful bone regeneration, as confirmed by histological analysis. Full article

Background and Objectives: The use of biomaterials in dentistry is extremely common. From a commercial perspective, different types of osteoconductive and osteoinductive biomaterials are available to clinicians. In the field of osteoconductive materials, clinicians have biomaterials made of heterologous bones at their disposal, including biomaterials of bovine, porcine, and equine origins, and biomaterials of natural origin, such as corals and hydroxyapatites. In recent years, it has become possible to synthesize nano-Ha and produce scaffolds using digital information. Although a large variety of biomaterials has been produced, there is no scientific evidence that proves their absolute indispensability in terms of the preservation of postextraction sites or in the execution of guided bone regeneration. While there is no scientific evidence showing that one material is better than another, there is evidence suggesting that several products have better in situ permanence. This article describes a preliminary study to evaluate the histological results, ISQ values, and prevalence of nano-HA. Materials and Methods: In this study, we planned to use a new biomaterial based on nanohydroxyapatite for implantation at one postextraction site; the nano-HA in this study was NuvaBONE (Overmed, Buccinasco, Milano, Italy). This is a synthetic bone graft substitute that is based on nanostructured biomimetic hydroxyapatite for application in oral–maxillofacial surgery, orthopedics, traumatology, spine surgery, and neurosurgery. In our pilot case, a patient with a hopeless tooth underwent extraction, and the large defect remaining after the removal of the tooth was filled with nano-HA to restore the volume. Twelve months later, the patient was booked for implant surgery to replace the missing tooth. At the time of the surgery, a biopsy of the regenerated tissue was taken using a trephine of 4 mm in the inner side and 8 mm deep. Results: The histological results of the biopsy showed abundant bone formation, high values of ISQ increasing from the insertion to the prosthetic phase, and a good reorganization of hydroxyapatite granules during resorption. The implant is in good function, and the replaced tooth shows good esthetics. Conclusions: The good results of this pilot case indicate starting the next Multicentric study to have more and clearer information about this nanohydroxyapatite (NH) compared with control sites. Full article

The stability of bone regenerated through Guided Bone Regeneration (GBR) around implants is crucial for long-term success. In this case series, changes in marginal bone levels (MBL) around implants placed in a regenerated bone using heterologous cortical lamina technique were radiographically measured. In addition, bone samples were obtained and submitted to histological and histomorphometric analysis. Thirty implants were placed in regenerated bone sites 8 months after the regenerative surgery; in the same surgical stage, a hard tissue biopsy was taken using a trephine bur and submitted to histologic and histomorphometric analysis. Changes in the marginal bone level, mesial and distal to the implant shoulder, were measured between prosthetic loading and the last follow-up, 2 years later. No implants were lost, and all could be deemed successful at the last follow-up. Only a minimal mean variation in the position of the marginal bone level was observed, both at the mesial (0.11 ± 0.49 mm) and at the distal level (0.03 ± 0.19 mm). The bone lamina had been resorbed after 8 months, and new bone had developed in close connection to the biomaterial. The average percentage of newly formed bone was 28%, while only 10% of the samples were composed of residual biomaterial; bone marrow and connective tissue composed the remaining part of the samples. This regeneration technique allowed, thanks to the rigidity of the lamina, the regeneration of new bone, which is stable after the prosthetic load. Further studies are needed to compare this procedure with those adopting non-resorbable, titanium-supported membranes. Full article

The diagnosis of systemic mastocytosis (SM) is based on various clinical, dermatological, serological, and hematological findings but essentially relies on histological evidence of an abnormal increase in tissue-localized mast cells (MCs). The extra-cutaneous organ most frequently affected is the bone marrow (BM), and therefore, histological examination of trephine biopsy specimens of the iliac crest is mandatory on suspicion of SM. At microscopic examination, neoplastic MCs show aberrant morphology, usually with prominent spindling. Immunohistochemistry is a useful tool in the diagnosis of SM because mast cell (MC) infiltrates may be slight and scarce, in a mixed background of lymphohistiocytic cells, eosinophils, and plasma cells. Moreover, neoplastic MCs exhibit an aberrant phenotype. Recent evidence, largely derived from molecular genetics, has enhanced the diagnostic capability of SM, also providing the basis for adequate prognostic and therapeutic evaluation. The cases herein reported illustrate the variable clinical manifestations and disease course of SM, focusing on diagnostic and therapeutic challenges. In accordance with the World Health Organization (WHO) classification and the International Consensus Classification (ICC) systems, our findings emphasize the importance of an integrated diagnostic approach for SM, with proper application of diverse assessment methodologies in order to improve SM classification and treatment effectiveness. Full article

(1) Background: The phenomenon of ankylosis of the dental elements has led clinicians to think that properly treated dentin and cement may be a potential graft for alveolar ridge augmentation. Currently, there are no studies in the literature able to histomorphometrically compare the healing patterns of an autogenous dentin particulate graft with the association, or not, of resorbable membranes. The aim of this pilot study is to histologically compare bone after an alveolar ridge augmentation using an autogenous dentin particulate graft with and without a resorbable collagen membrane. (2) Methods: this clinical trial enrolled six patients with defects requiring bone augmentation. Two procedures were performed in all six adult human patients in order to perform a study–control study: in Group 1, a ridge augmentation procedure with an autogenous dentin particulate graft and a resorbable collagen membrane was performed, and, in Group 2, an alveolar ridge preservation without a membrane was performed at the same time (T0). At 4 months, a biopsy of the bone tissues was performed using a 4 mm trephine bur in order to perform a histomorphometric analysis. (3) Results: The histomorphometric analysis demonstrated that Group 1 presented 45% of bone volume, 38% of vital bone, and 7% of residual graft. On the contrary, membrane-free regenerative procedures demonstrated 37% of bone volume, 9% of vital bone, and 27% of non-resorbed graft. In all cases, the regenerated bone allowed the insertion of implants with a standard platform, and no early failures were recorded. (4) Conclusions: Autogenous dentin particulate grafts seem to work best when paired with a membrane. Full article

Mesenchymal stromal cells (MSC) ‘educated’ by tumor cells are an essential component of the multiple myeloma (MM) tumor microenvironment (TME) involved in tumor progression. Transcription of tandemly repeated (TR) non-coding DNA is often activated in many tumors and is required for tumor progression and cancer cells genome reorganization. The aim of the work was to study functional properties including the TR DNA transcription profile of MSC from the hematopoietic niche of treated MM patients. Healthy donors (HD) and patients after bortezomib-based treatment (with partial or complete response, PoCR, and non-responders, NR) were enrolled in the study. Their trephine biopsies were examined histologically to evaluate the hematopoietic niche. MSC cultures obtained from the biopsies were used for evaluation of the proliferation rate, osteogenic differentiation, presence of tumor MSC markers, resistance to bortezomib, and pericentromeric TR DNA transcription level. The MSC ‘education’ by multiple myeloma cells was mimicked in co-culture experiments with or without bortezomib. The TR DNA transcription profile was accessed. The histological examination revealed the persistence of the tumor microenvironment (especially of the vasculature) in treated patients. In co-culture experiments, MSC of bortezomib-treated patients were more resistant to bortezomib and protected cancer MM cells of the RPMI8226 cell line more effectively than HD-MSC did. The MSC obtained from PoCR and NR samples differed in their functional properties (proliferation capacity, osteogenic potential, and cancer-associated fibroblasts markers). Transcriptome analysis revealed activation of the TR transcription in cells of non-hematopoietic origin from NR patients’ bone marrow. The pericentromeric TR DNA of HS2/HS3 families was among the most upregulated in stromal MSC but not in cancer cells. The highest level of transcription was observed in NR-MSC. Transcription of HS2/HS3 was not detected in healthy donors MSC unless they were co-cultured with MM cancer cells and acquired cancer-associated phenotype. Treatment with TNFα downregulated HS2/HS3 transcription in MSC and upregulated in MM cells. Our results suggest that the hematopoietic niche retains the cancer-associated phenotype after treatment. Pericentromeric non-coding DNA transcription is associated with the MSC cancer-associated phenotype in patients with ineffective or partially effective multiple myeloma treatment. Full article

Aim: The aim of this technical note is to present a computer-aided design–computer-aided manufacturing (CAD–CAM) surgical guide to perform a computer-guided bone biopsy. Traditionally, to diagnose abnormal conditions affecting jawbone, a bone biopsy is performed with the use of a trephine bur. The positioning of the bur, during the biopsy, is based on the skill of the surgeon; therefore, an inaccurate placement of a trephine bur may occur. The use of a guide, however, can minimize this risk and achieve a better result. Materials and Methods: To determine the site and the extension of bone sampling, the stereolithography file (STL) file of cone–beam computed tomography (CBCT) images is acquired using a specific planning software and superimposed with the STL file of a dental cast; a virtual surgical guide is designed, using the same software that allows a 3D (three-dimensional) view of the guide from different perspectives and planes. The number and site of guide tubes are determined on the basis of the width and the extension of the sampling; thanks to a 3D printer, the surgical guide is manufactured. Results: The use of a customized surgical guide realized with CAD–CAM technology allows a precise and minimally invasive approach, with an accurate three-dimensional localization of the biopsy site. Conclusions: The high precision, great predictability, time-effectiveness and versatility of the present guide should encourage the clinician to use this minimally invasive surgical approach, but controlled clinical trials should be conducted to evaluate the advantages as well as any possible complications. Full article

The diagnosis of a myeloid neoplasm relies on a combination of clinical, morphological, immunophenotypic and genetic features, and an integrated, multimodality approach is needed for precise classification. The basic diagnostics of myeloid neoplasms still rely on cell counts and morphology of peripheral blood and bone marrow aspirate, flow cytometry, cytogenetics and bone marrow trephine biopsy, but particularly in the setting of Ph− myeloproliferative neoplasms (MPN), the trephine biopsy has a crucial role. Nowadays, molecular studies are of great importance in confirming or refining a diagnosis and providing prognostic information. All myeloid neoplasms of chronic evolution included in this review, nowadays feature the presence or absence of specific genetic markers in their diagnostic criteria according to the current WHO classification, underlining the importance of molecular studies. Crucial differential diagnoses of Ph− MPN are the category of myeloid/lymphoid neoplasms with eosinophilia and gene rearrangement of PDGFRA, PDGFRB or FGFR1, or with PCM1-JAK2, and myelodysplastic/myeloproliferative neoplasms (MDS/MPN). This review focuses on morphological, immunophenotypical and molecular features of BCR-ABL1-negative MPN and their differential diagnoses. Furthermore, areas of difficulties and open questions in their classification are addressed, and the persistent role of morphology in the area of molecular medicine is discussed. Full article

The aim of the present clinical study was to assess and compare the histomorphometric results and efficacy of freeze-dried bone allograft (FDBA) in combination with platelet-rich fibrin (PRF), and PRF as a sole grafting material for socket preservation. Ninety patients in need of tooth extraction and implant restoration were included in this study. The participants were randomly divided into three groups based on post-extraction clinical protocol: socket preservation procedure with allograft in combination with a PRF membrane (PRFm), PRF as a sole grafting material, and a control group. A total of 90 implants were placed four months post-extraction. During the surgical re-entry a bone biopsy was harvested with a trephine drill. Histological samples were prepared and analyzed for percentage vital bone and connective tissue. One-way ANOVA with Bonferroni post-hoc analysis were used to assess the results. Both test groups revealed a significantly higher percentage of vital bone formation compared to the control group. No statistically significant differences regarding vital bone formation and connective tissue quantity between the tested groups were observed (FDBA + PRFm: 3.29 ± 13.03%; and PRF: 60.79 ± 9.72%). From a clinical and histological point of view, both materials in the test groups are suitable for the filling of post-extraction sockets without bone defects. Both of the tested groups revealed a significantly higher percentage of vital bone formation compared to the control group. Full article

Background: This study aims to evaluate whether there is a correlation between implant stability, bone density, vital bone formation and implant diameter and length. Methods: Ninety patients were enrolled in this study. They underwent a socket preservation procedure with allograft or PRF and after 4 months, a total of 90 implants were placed. CBCT scans were assigned prior to implant placement in order to assess the bone density. During the surgical re-entry, a bone biopsy was harvested with a trephine drill. Immediately after implant insertion, the primary stability was measured. The secondary stability was measured 4 months after implant placement. Results: Primary stability showed a significant positive linear correlation with bone density (r = 0.471, p < 0.001) as well as with percentage of new bone formation (r = 0.567, p < 0.001). An average significant association of secondary stability with bone density (rs = 0.498, p < 0.001) and percentage of newly formed bone (r = 0.477, p < 0.001) was revealed. The mean values of primary stability in all three implant sizes, regarding the diameter of the implants, were similar (narrow 67.75; standard 66.78; wide 71.21) with no significant difference (p = 0.262). The same tendency was observed for secondary stability (narrow 73.83; standard 75.25; wide 74.93), with no significant difference (p = 0.277). Conclusions: The study revealed a high correlation between primary and secondary implant stability, and bone density, as well as with the percentage of vital bone formation. Implant length and diameter revealed no linear correlation with the implant stability. Full article

Aim: To evaluate the hypothesis of a correlation between the preoperative residual alveolar bone height (RBH) and graft maturation after maxillary sinus floor augmentation procedures using two different bone substitutes. Methods: A total of 20 patients who underwent unilateral maxillary sinus floor augmentation with either mineralized deproteinized bovine bone (DBBM) or a xenograft enriched with polymer and gelatin (NBS) were included in this prospective study. Six months after sinus surgery, bone biopsies were harvested with a 3.2 mm diameter trephine bur, prior to dental implant placement. Histomorphometric analysis was performed, and the results were correlated with the individual RBH. Implants were loaded after 5 months of insertion, and 1-year implant success and marginal bone level change were assessed. Results: RBH was 2.17 ± 1.11 mm (range 0.5–3.5 mm) and 2.14 ± 0.72 mm (range 0.5–3.0 mm) in the NBS and DBBM group, respectively. The biopsy analyses for the DBBM group showed woven bone increases by 5.08% per 1-mm increment of RBH; medullary spaces decreased by 9.02%, osteoid decreased by 4.4%, residual biomaterial decreased by 0.34%, and lamellar bone increased by 5.68% per 1-mm increase of RBH. In the NBS group, samples showed woven bone increases by 8.08% per 1-mm increase of RBH; medullary spaces decreased by 0.38%; osteoid increased by 1.34%, residual biomaterial decreased by 0.58%, and lamellar bone decreased by 5.50% per 1-mm increase of RBH. There was no statistically significant difference in the correlation between RBH and lamellar bone, woven bone, and osteoid, independently of the material used. Implant success was 100% in both groups, and marginal bone loss was 1.02 ± 0.42 mm in DBBM and 0.95 ± 0.31 mm in the NBS group after the 1-year follow-up. Conclusion: In spite of the absence of significance, the observed trend for woven bone to increase and medullary spaces to decrease when RBH increases deserves attention. Residual bone dimension might be a determinant in the bone graft maturation after maxillary sinus augmentation. Full article

This randomized controlled clinical trial evaluated the effect of mineralized plasmatic matrix (MPM), comprised of synthetic graft and platelet concentrates, on new bone formation and volume stability over time in maxillary sinus lifting (MSL). Unilateral MSL was performed in 20 patients with either beta-tricalcium phosphate (β-TCP) or MPM grafts (10 sinuses each). Six months postsurgery, specimens were obtained with a trephine bur prior to implant placement in 39 cases. Volumetric changes in sinus augmentation were analyzed between 1 week (T-I) and 6 months (T-II) postsurgery. Histomorphometric and histological analyses of biopsy samples revealed mean new bone percentages of 35.40% ± 9.09% and 26.92% ± 7.26% and residual graft particle areas of 23.13% ± 6.16% and 32.25% ± 8.48% in the MPM and β-TCP groups, respectively (p < 0.05). The mean soft-tissue areas in the MPM and β-TCP groups were 41.48% ± 8.41% and 40.83% ± 8.86%, respectively (p > 0.05). Graft reductions between baseline and 6-months postprocedure in the β-TCP and MPM groups were 17.12% ± 13.55% and 14.41% ± 12.87%, respectively, with significant graft volume reduction observed in both groups (p < 0.05) while there is no significant difference between MPM and β-TCP groups (p > 0.05). Thus, MPM, representing growth factors in a fibrin network, increases new bone formation and has acceptable volume stability in MSL procedures Full article