Polylevolysine and Fibronectin-Loaded Nano-Hydroxyapatite/PGLA/Dextran-Based Scaffolds for Improving Bone Regeneration: A Histomorphometric in Animal Study.

The regeneration of large bone defects is still demanding, requiring biocompatible scaffolds, with osteoconductive and osteoinductive properties. This study aimed to assess the pre-clinical efficacy of a nano-hydroxyapatite (nano-HA)/PGLA/dextran-based scaffold loaded with Polylevolysine (PLL) and fibronectin (FN), intended for bone regeneration of a critical-size tibial defect, using an ovine model. After physicochemical characterization, the scaffolds were implanted in vivo, producing two monocortical defects on both tibiae of ten adult sheep, randomly divided into two groups to be euthanized at three and six months after surgery. The proximal left and right defects were filled, respectively, with the test scaffold (nano-HA/PGLA/dextran-based scaffold loaded with PLL and FN) and the control scaffold (nano-HA/PGLA/dextran-based scaffold not loaded with PLL and FN); the distal defects were considered negative control sites, not receiving any scaffold. Histological and histomorphometric analyses were performed to quantify the bone ingrowth and residual material 3 and 6 months after surgery. In both scaffolds, the morphological analyses, at the SEM, revealed the presence of submicrometric crystals on the surfaces and within the scaffolds, while optical microscopy showed a macroscopic 3D porous architecture. XRD confirmed the presence of nano-HA with a high level of crystallinity degree. At the histological and histomorphometric evaluation, new bone formation and residual biomaterial were detectable inside the defects 3 months after intervention, without differences between the scaffolds. At 6 months, the regenerated bone was significantly higher in the defects filled with the test scaffold (loaded with PLL and FN) than in those filled with the control scaffold, while the residual material was higher in correspondence to the control scaffold. Nano-HA/PGLA/dextran-based scaffolds loaded with PLL and FN appear promising in promoting bone regeneration in critical-size defects, showing balanced regenerative and resorbable properties to support new bone deposition.

Tamoxifen improves homeostasis in the peri-implant bone remodeling of osseointegrated titanium implants.

BACKGROUND: The purpose of this preclinical study was to evaluate the influence of tamoxifen (TAM) on the peri-implant bone remodeling of osseointegrated titanium implants in ovariectomized female rats. MATERIALS AND METHODS: Seventy-two female rats underwent bilateral ovariectomy 20 weeks before implants placement. One titanium implant was inserted in each tibia of the animals. Six weeks following the implant surgery, animals were randomly divided into two experimental groups (n = 36), which received either saline solution (SS) or tamoxifen citrate (TAM) via gavage until euthanasia. Euthanasia was performed at 30, 60, and 90 days after the first gavage. Assessments of bone to implant contact (BIC), bone ingrowth percentage (BIN), morphological description of cellular and tissue reactions, immunohistochemistry for the detection of bone morphogenetic protein 2/4 (BMP2/4), runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN) and tartrate-resistant acid phosphatase (TRAP), and bone chemical composition through scanning electron microscopy with energy-dispersive x-ray spectroscopy were performed. RESULTS: Tamoxifen group presented higher BIC, higher BIN, higher RUNX-2 and OCN, lower TRAP-positive cells/mm2 , and no differences regarding BMP-2/4 positive cells/mm2 than SS group in all periods. TAM group also showed higher Ca/P rate than SS group. CONCLUSION: Tamoxifen enhanced the remodeling of the bone surrounding titanium implants in ovariectomized rats.

Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study.

Background and Objectives: Three-dimensional (3D) metallic trabecular structures made by additive manufacturing (AM) technologies promote new bone formation and osteointegration. Surface modifications by chemical treatments can improve the osteoconductive properties of metallic structures. An in vivo study in sheep was conducted to assess the bone response to randomized trabecular titanium structures that underwent a surface modification by chemical treatment compared to the bone response to the untreated specimens. Material and Methods: Sixteen specimens with a randomized trabecular titanium structure were implanted in the spongious bone of the distal femur and proximal tibia and the cortical bone of the tibial diaphysis of two sheep. Of them, eight implants had undergone a chemical treatment (treated) and were compared to eight implants with the same structure but native surfaces (native). The sheep were sacrificed at 6 weeks. Surface features of the lattice structures (native and treated) were analyzed using a 3D non-contact profilometer. Compression tests of 18 lattice cubes were performed to investigate the mechanical properties of the two structures. Excellent biocompatibility for the trabecular structures was demonstrated in vitro using a cell mouse fibroblast culture. Histomorphometric analysis was performed to evaluate bone implant contact and bone ingrowth. Results: A compression test of lattice cubic specimens revealed a comparable maximum compressive strength value between the two tested groups (5099 N for native surfaces; 5558 N for treated surfaces; p > 0.05). Compared to native surfaces, a homogenous formation of micropores was observed on the surface of most trabeculae that increased the surface roughness of the treated specimens (4.3 versus 3.2 µm). The cellular viability of cells seeded on three-dimensional structure surfaces increased over time compared to that on plastic surfaces. The histomorphometric data revealed a similar behavior and response in spongious and cortical bone formation. The percentage of the implant surface in direct contact with the regenerated bone matrix (BIC) was not significantly different between the two groups either in the spongious bone (BIC: 27% for treated specimens versus 30% for native samples) or in the cortical bone (BIC: 75% for treated specimens versus 77% for native samples). Conclusions: The results of this study reveal rapid osseointegration and excellent biocompatibility for the trabecular structure regardless of surface treatment using AM technologies. The application of implant surfaces can be optimized to achieve a strong press-fit and stability, overcoming the demand for additional chemical surface treatments.

Effects of Vitamin and Amino Acid-Enriched Hyaluronic Acid Gel on the Healing of Oral Mucosa: In Vivo and In Vitro Study.

Background and Objectives: Wound healing is a dynamic process that can be compromised in patients with chronic and metabolic conditions or unhealthy lifestyles. Numerous medical substances designed for topical use, charged with compounds that promote the healing process, have been developed to improve wound healing, especially in compromised subjects. The present study aimed to extend our understanding of the in vivo effects of a hyaluronic acid gel charged with amino acids (HAplus gel, Aminogam gel® Errekappa Euroterapici spa, Milan, Italy) and study the in vitro effects of the same gel charged with additional substances in an attempt to optimize its formulation. Materials and Methods: In a randomized controlled split-mouth clinical and histological trial, HAplus gel was tested on the gingival tissue of the lower third molar post-extraction socket. The gingiva was collected at the time of extraction (T0) and ten days after the extraction (T1) to be histologically analyzed. During the second stage of the study, culture media with HAplus gel and vitamin C and E at different concentrations (TEST) were tested on human gingival fibroblasts and compared to the HAplus-enriched medium (HA-Control). Results: Histological and immunohistochemical analysis of collected gingiva showed higher microvascular density and collagen fibers organized in closely packed and well-oriented bundles in sites treated with HAplus gel. In the in vitro study, all TEST groups showed an increased viability from 24 h to 48 h. After 24 h, the viability percentage in all experimental groups was below 100% of the HA-Control, demonstrating a mild toxicity. After 48 h from seeding, the TEST groups' viability grew significantly compared to HA-Control. Conclusions: These encouraging preliminary results suggest that the use of HAplus gel enriched with vitamins C and E may be beneficial in patients with conditions that impair soft tissue healing.

In vivo osseointegration of a randomized trabecular titanium structure obtained by an additive manufacturing technique.

The additive manufacturing techniques (AM) are able to realize three-dimensional trabecular structures that mimic the trabecular structure of the bone. An in vivo study in sheep was carried out with the aim of assessing the bone response and the trend of osteointegration of a randomized trabecular titanium structure produced by the AM technique. In 6 sheep were implanted 84 specimens with a trabecular titanium structure (4 implants in the femur distal epiphysis; 4 implants in the tibial plate; 6 implants in the tibial shaft). Sheep were sacrificed at 3 postoperative time-points: 6 weeks, 10 weeks, 14 weeks. Histomorphometric analysis was performed for the evaluation of Bone Implant Contact, and Bone Ingrowth. A standard push-out test was used to analyze the mechanical characteristics of the bone-implant interface. The histomorphometric data and biomechanical tests showed a fast osseointegration of the specimens both in the cancellous and in the cortical bone. The quantitative analysis of osseointegration data in cancellous bone showed the percentage of the surface of the implant in direct contact with the regenerated bone matrix significantly improved from 28% at 6 weeks to 54% at 14 weeks. An early osseointegration occurred in cortical bone showing that 75% of surface of implant was in direct contact with regenerated bone after 6 weeks; this value increased to 85% after 14 weeks. Mechanical tests revealed an early improvement of mean peak load of implants at 10 weeks (4486 N ± 528 N) compared to values at 6 weeks (2516 N ± 910 N) confirming the high rate of progression of osseointegration in the cortical bone. The non-mineralized matrix followed an increasing process of mineralization almost completely after 14 weeks. The results of this study have showed a rapid osseointegration and excellent biocompatibility for a randomized trabecular titanium structure that should be confirmed by clinical investigations.

Ridge Preservation Using a Novel Freeze-dried Enzyme-deantigenic Bone Paste: A Histomorphometric-retrospective Pilot Case Series.

AIM AND OBJECTIVE: The aim of this study is to provide preliminary retrospective evidence concerning the histologic and histomorphometric outcome of a novel freeze-dried equine-derived bone paste (EDEBEX) for ridge preservation of sockets following tooth extraction. MATERIALS AND METHODS: This pilot retrospective case series describes the histologic and histomorphometric outcome of three patients who received the equine-derived bone paste in post-extractive sockets to allow the preservation of the alveolar ridge. Patients were later rehabilitated with monolithic-zirconia, implant-supported prostheses. RESULTS: All patients healed uneventfully. The collected biopsies showed a prevalence of bone formation at 4 months, compact lamellar bone, with well-defined lamellae surrounding Haversian and Volkmann's canals at 6 months, and an intermediate degree of maturation in active anabolic phase at 7 months after grafting. The amount of mineralized matrix was 63.3-70.7%, whereas medullar spaces were 26.0-30.7%. CONCLUSION: Histologic examination showed that the bone paste was fully biocompatible. Bone regeneration occurred within the first 4 months from grafting, with 63.3-70.7% mineralized bone matrix. The residual biomaterial, when present, did not exceed, on average, 2%. CLINICAL SIGNIFICANCE: Ridge preservation using bone substitutes as an alternative to autogenous bone is known to be effective. However, available clinical evidence still does not indicate the biomaterial, if any, that should be preferred to carry it out. The equine bone paste used in the present study appears to be a good candidate for further investigation because it is easy to handle in the clinical setting and it displays a good bone formation rate.

Correlational study on altered epicardial adipose tissue as a stratification risk factor for valve disease progression through IL-13 signaling.

AIMS: Genetic and environmental factors all interact in the risk of progression of valvular dysfunctions. Previous studies reported a relation between valve diseases and epicardial adipose tissue (EAT) thickness. The aim of this study was to verify the possible relationship between the molecular pattern of EAT related to IL-13 fibrogenic cytokine expression and valve dysfunction. METHODS AND RESULTS: A valvular heart disease (VHD) population was stratified according to their median EAT thickness (7 mm). The molecular expression of IL-13 in EAT is directly related to the molecular expression of genes associated with extracellular matrix (ECM) turnover, macrophage infiltration and promotion of the formation of ectopic calcific nodules involved in aorta coarctation and calcification. CONCLUSION: IL-13 gene expression in altered EAT is directly related to the expression of genes involved in ECM turnover and the formation of ectopic calcific nodules, suggesting measurements of EAT as a stratification risk factor for valve instability in the VHD patients.

CoCrMo alloys ions release behavior by TiNbN coating: an in vitro study.

The aim of the study was to show in vitro the greater inertness to the corrosion body fluid of TiNbN coating than the CoCrMo alloy substrate. The prosthetic component under study was a femoral component of total knee prosthesis in CoCrMo alloy coated in TiNbN with Physical Vapor Deposition technique immersed in static Hank's balanced salt solution (HBS) (pH = 6) for at least 34 months at a constant temperature of 37 °C. Another uncoated prosthetic component of CoCrMo alloy with the same type and size was left in static immersion in the same solution and for the same period of time. Scanning electron microscope (SEM) analysis was performed to investigate adhesion and proliferation at 24, 48, 72 h after seeding of 104 sub-confluents osteoblast-like cells (SaOS-2) cells on scaffold. The results of the study showed a reduction in the concentration of the metal ions released from the TiNbN-coated femoral component surface compared to the uncoated surface in the HBS solution. The overall reduction of the ions for the TiNbN-coated femoral component compared to the uncoated one was 80.1 ± 2%, 62.5% ± 8% and 48% ± 10% for Co, Cr, Mo, respectively (p < 0.01). SEM analysis confirmed the healthy state of the cells, the cellular adhesion and proliferation of SaOS-2 on the TiNbN-coated specimen. Although the results observed in vitro for the TiNbN coating are encouraging, clinical studies are certainly needed to be performed in order to understand how these positive findings can be translated in vivo and to determine the clinical benefit of TiNbN coating.

CEMP-1 Levels in Periodontal Wound Fluid during the Early Phase of Healing: Prospective Clinical Trial.

OBJECTIVES: Cementogenesis seems to be significantly compromised during tissue inflammation. In dental practice, surgical procedures are performed with the aim to regenerate periodontium including cementum. However, inflammation that occurs during the initial healing phases after surgery may impair regeneration of this tissues. The aim of the present study was to assess if surgical procedures designed to regenerate periodontium might affect levels of cementum protein-1 (CEMP-1) in periodontal wound fluid during early phase of healing. MATERIALS AND METHODS: In 36 patients, 18 intrabony periodontal defects were treated with regenerative therapy (REG group) and 18 suprabony periodontal defects were treated with open flap debridement (OFD group). In the experimental sites, gingival crevicular fluid was collected immediately before surgery, and periodontal wound fluid was collected 4, 7, 14, and 21 days after surgery. CEMP-1 levels were detected by indirect enzyme-linked immunosorbent assay technique. RESULTS: At the analysis, it resulted that there was a significant average difference in CEMP-1 values between the REG and OFD groups at baseline (p = 0.041), the CEMP-1-modeled average in the OFD group was lower by 0.45 ng/ml. There was a significant trend in CEMP-1 over time, and this trend was different among the 2 groups: the REG group showed a statistically significant rising CEMP-1 trend (0.18 ng/ml a week p = 0.012), while the OFD had a trend that was significantly lower (-0.22 ng/ml a week compared to the REG group trend p = 0.023), the OFD group lost on average 0.05 ng/ml a week. In REG sites, GCF protein levels resulted also related to clinical parameters. CONCLUSIONS: During the initial inflammatory phase of periodontal healing, CEMP-1 levels decrease regardless of the surgical protocol applied. The surgical procedures used to regenerate periodontal tissue are able to reverse this trend and to induce significant increase of CEMP-1 in periodontal wound fluid after the first week postop.

Osseointegration of Titanium Implants With Different Rough Surfaces: A Histologic and Histomorphometric Study in an Adult Minipig Model.

BACKGROUND: Many chemical and physical modifications of titanium surfaces were introduced, aiming at improving surface bioactivity, but few comparative evidence exists. OBJECTIVE: To evaluate histologically in minipigs the osseointegration of implants made of commercially pure (CP) titanium or titanium alloy, treated by different roughening procedures. MATERIAL AND METHODS: Three sandblasted acid-etched (SA) surfaces, 2 anodized (AN), and 1 double acid-etched (DAE) were compared. Surface microtopography was characterized with scanning electron microscope; surface element composition was also assessed. One implant per group was inserted in each proximal tibia of 2 minipigs. Three months after healing, block biopsies were taken for histomorphometric analysis. Implant stability quotient (ISQ) was measured at insertion and before harvesting. RESULTS: The highest amount of cortical bone-implant contact was observed around SA implants and showed positive correlation with surface roughness. The greatest increase in ISQ was observed in CP-AN implants. In the medullary region, SA implants showed the best osteogenic response, whereas inflammatory cells were found around DAE and alloy-AN implants. CONCLUSIONS: SA surfaces were more osteogenic than anodized or dual acid-etched ones, although not significantly. Surface roughness affected osseointegration.

Human Adipose-Derived Stem Cells on Rapid Prototyped Three-Dimensional Hydroxyapatite/Beta-Tricalcium Phosphate Scaffold.

In the study, we assess a rapid prototyped scaffold composed of 30/70 hydroxyapatite (HA) and beta-tricalcium-phosphate (ß-TCP) loaded with human adipose-derived stem cells (hASCs) to determine cell proliferation, differentiation toward osteogenic lineage, adhesion and penetration on/into the scaffold.In this in vitro study, hASCs isolated from fat tissue discarded after plastic surgery were expanded, characterized, and then loaded onto the scaffold. Cells were tested for: viability assay (Alamar Blue at days 3, 7 and Live/Dead at day 32), differentiation index (alkaline phosphatase activity at day 14), scaffold adhesion (standard error of the mean analysis at days 5 and 18), and penetration (ground sections at day 32).All the hASC populations displayed stemness markers and the ability to differentiate toward adipogenic and osteogenic lineages.Cellular vitality increased between 3 and 7 days, and no inhibitory effect by HA/ß-TCP was observed. Under osteogenic stimuli, scaffold increased alkaline phosphatase activity of +243% compared with undifferentiated samples. Human adipose-derived stem cells adhered on HA/ß-TCP surface through citoplasmatic extensions that occupied the macropores and built networks among them. Human adipose derived stem cells were observed in the core of HA/ß-TCP. The current combination of hASCs and HA/ß-TCP scaffold provided encouraging results. If authors' data will be confirmed in preclinical models, the present engineering approach could represent an interesting tool in treating large bone defects.

Iliac Crest Fresh-Frozen Allografts Versus Autografts in Oral Pre-Prosthetic Bone Reconstructive Surgery: Histologic and Histomorphometric Study.

PURPOSE: The aim of this work was to compare histologically bone biopsies obtained during dental implant placement in atrophic jaws reconstructed with fresh-frozen bone (FFB) or autologous bone blocks. MATERIALS AND METHODS: Twenty partially or totally edentulous patients were treated either with iliac crest FFB grafts (14 patients; group A) or with iliac crest autografts (6 patients; group B). Seven to 9 months later, dental implants were placed in the reconstructed jaws and bone biopsies were harvested. RESULTS: The reconstructive procedure was uneventful in 9 patients of group A and in all patients of group B. Samples from both groups did not show relevant differences concerning the proportion of lamellar bone, newly formed bone, and bone marrow. In group A, the decreasing proportion of blood vessels from the apical to the coronal portion, the larger amount of lacunae in the apical portion, and the fewer vessels in the coronal portion than in group B evoke an effort of the host bone to "integrate" the graft from the apical portion. CONCLUSIONS: FFB cannot be considered as successful and safe in alveolar bone reconstruction as autogenous bone grafting.

Osseointegration of implants with dendrimers surface characteristics installed conventionally or with Piezosurgery®. A comparative study in the dog.

AIM: The first aim of the present experiment was to compare bone healing at implants installed in recipient sites prepared with conventional drills or a piezoelectric device. The second aim was to compare implant osseointegration onto surfaces with and without dendrimers coatings. MATERIAL AND METHODS: Six Beagles dogs were used in this study. Five implants with two different surfaces, three with a ZirTi(®) surface (zirconia sand blasted, acid etched), and two with a ZirTi(®)-modified surface with dendrimers of phosphoserine and polylysine were installed in the right side of the mandible. In the most anterior region (P2, P3), two recipient sites were prepared with drills, and one implant ZirTi(®) surface and one coated with dendrimers implants were installed at random. In the posterior region (P4 and M1), three recipient sites were randomly prepared: two sites with a Piezosurgery(®) instrument and one site with drill and two ZirTi(®) surface and one coated with dendrimers implants installed. Three months after the surgery, the animals were sacrificed for histological analysis. RESULTS: No complications occurred during the healing period. Three implants were found not integrated and were excluded from analysis. However, n = 6 was obtained. The distance IS-B at the buccal aspect was 2.2 ± 0.8 and 1.8 ± 0.5 mm, while IS-C was 1.5 ± 0.9 and 1.4 ± 0.6 mm at the Piezosurgery(®) and drill groups, respectively. Similar values were obtained between the dendrimers-coated and ZirTi(®) surface implants. The BIC% values were higher at the drill (72%) compared to the Piezosurgery(®) (67%) sites. The BIC% were also found to be higher at the ZirTi(®) (74%) compared to the dendrimers-coated (65%) implants, the difference being statistically significant. CONCLUSION: This study has revealed that oral implants may osseointegrate equally well irrespective of whether their bed was prepared utilizing conventional drills with abundant cooling or Piezosurgery(®). Moreover, the surface coating of implants with dendrimers phosphoserine and polylysine did not improve osseointegration.

Focal Adhesion's Role in Cardiomyocytes Function: From Cardiomyogenesis to Mechanotransduction.

Mechanotransduction refers to the ability of cells to sense mechanical stimuli and convert them into biochemical signals. In this context, the key players are focal adhesions (FAs): multiprotein complexes that link intracellular actin bundles and the extracellular matrix (ECM). FAs are involved in cellular adhesion, growth, differentiation, gene expression, migration, communication, force transmission, and contractility. Focal adhesion signaling molecules, including Focal Adhesion Kinase (FAK), integrins, vinculin, and paxillin, also play pivotal roles in cardiomyogenesis, impacting cell proliferation and heart tube looping. In fact, cardiomyocytes sense ECM stiffness through integrins, modulating signaling pathways like PI3K/AKT and Wnt/ß-catenin. Moreover, FAK/Src complex activation mediates cardiac hypertrophic growth and survival signaling in response to mechanical loads. This review provides an overview of the molecular and mechanical mechanisms underlying the crosstalk between FAs and cardiac differentiation, as well as the role of FA-mediated mechanotransduction in guiding cardiac muscle responses to mechanical stimuli.

Empty vein ablation (EVA) technique: an in-vivo animal model to assess the effects of sclerosing agent concentration and wall contact time on intima and media tunicae structure.

BACKGROUND: Sclerotherapy is a cornerstone of the treatment of chronic venous disease, despite some technical aspects (e.g., sclerosant liquid agent concentration [SLAC] and contact time between sclerosant agent and vein wall [ctSA/VW]) to maximize outcomes remain an unsolved problem and a source of debate. An innovative three-balloon catheter has been developed to allow sclerotherapy in empty vein conditions (Empty Vein Ablation technique, EVA), revolutionizing the definition of SLAC and ctSA/VW. Aim of this experimental study is to analyze EVA effects on intima and media vessel tunicae using different SLAC and ctSA/VW in an in-vivo animal model. METHODS: Two adult sheep were treated by EVA using jugular and common iliac vein axes (eight vein segments). Different SLAC (polidocanol 0.5% or 1%) and different ctSA/VW (3 or 5 minutes) were combined for testing residual circumferential intima percentage and media thickness after EVA. RESULTS: Intact circumferential residual intima after the treatment was 21.3±4.9%, 18.2±7.4%, 15.7±2.4% and 8.9±2.0% using 0.5% (3 min), 0.5% (5 min), 1% (3 min) and 1% (5 min), respectively (R2=0.945; control sample: 97.6%). Media thickness after the treatment was 121.6±35.3 µm, 110.9±7.8 µm, 96.1±30.4 µm and 79.1±34.1 µm using 0.5% (3 min), 0.5% (5 min), 1% (3 min) and 1% (5 min), respectively (R2=0.990; control sample 125.7 µm). No significant modifications were detected analyzing the adventitia in all samples. CONCLUSIONS: EVA proved to be effective in venous wall destruction even with a very low SLAC and ctSA/VW (0.5% in 3 minutes), in quite large caliber veins. Direct comparisons with foam/liquid sclerotherapy should be done to confirm therapeutic effectiveness of these results, despite EVA has provided a maximized and controlled SA/VW contact time and ratio.

Histological findings on jaw osteonecrosis associated with bisphosphonates (BONJ) or with radiotherapy (ORN) in humans.

OBJECTIVE: To describe the histological features of bone tissue harvested from patients affected by jaw osteonecrosis associated with bisphoshponates (BONJ) or with radiotherapy (ORN), in undecalcified ground sections. MATERIALS AND METHODS: Sixteen bone tissue samples from 14 patients with BONJ and two patients with ORN were processed in order to obtain both ground, undecalcified sections and decalcified sections. The sections underwent histometric and morphometric analysis. RESULTS: Bone tissue samples obtained from patients with BONJ or ORN of the jaws shared some histological characteristics. Common histological features included the loss of bone architecture, the absence of a proper Haversian system and proper marrow spaces, the presence of necrotic spots of non-mineralized tissue, areas of empty osteocytic lacunae next to areas of hypercellularity, the presence of resorption pits with rare osteoclast-like cells and the presence of bacteria and of an inflammatory infiltrate. A violet rib of tissue characterized by large resorption pits facing was frequently observed between the mineralized bone and the inflammatory infiltrate. CONCLUSIONS: The histological features of BONJ and ORN are similar and resemble those of osteomyelitis. Even though it is not clear whether infection is the cause or consequence of bone exposure, inflammatory cells, bacteria or their products may have a massive, direct lytic effect on bone tissue challenged by bisphosphonates.

Cell-Laden 3D Printed GelMA/HAp and THA Hydrogel Bioinks: Development of Osteochondral Tissue-like Bioinks.

Osteochondral (OC) disorders such as osteoarthritis (OA) damage joint cartilage and subchondral bone tissue. To understand the disease, facilitate drug screening, and advance therapeutic development, in vitro models of OC tissue are essential. This study aims to create a bioprinted OC miniature construct that replicates the cartilage and bone compartments. For this purpose, two hydrogels were selected: one composed of gelatin methacrylate (GelMA) blended with nanosized hydroxyapatite (nHAp) and the other consisting of tyramine-modified hyaluronic acid (THA) to mimic bone and cartilage tissue, respectively. We characterized these hydrogels using rheological testing and assessed their cytotoxicity with live-dead assays. Subsequently, human osteoblasts (hOBs) were encapsulated in GelMA-nHAp, while micropellet chondrocytes were incorporated into THA hydrogels for bioprinting the osteochondral construct. After one week of culture, successful OC tissue generation was confirmed through RT-PCR and histology. Notably, GelMA/nHAp hydrogels exhibited a significantly higher storage modulus (G') compared to GelMA alone. Rheological temperature sweeps and printing tests determined an optimal printing temperature of 20 °C, which remained unaffected by the addition of nHAp. Cell encapsulation did not alter the storage modulus, as demonstrated by amplitude sweep tests, in either GelMA/nHAp or THA hydrogels. Cell viability assays using Ca-AM and EthD-1 staining revealed high cell viability in both GelMA/nHAp and THA hydrogels. Furthermore, RT-PCR and histological analysis confirmed the maintenance of osteogenic and chondrogenic properties in GelMA/nHAp and THA hydrogels, respectively. In conclusion, we have developed GelMA-nHAp and THA hydrogels to simulate bone and cartilage components, optimized 3D printing parameters, and ensured cell viability for bioprinting OC constructs.

Empty vein ablation innovative technique for chronic venous disease treatment: proof of concept and ex-vivo analysis.

BACKGROUND: Sclerotherapy is among the mainstays of chronic venous disease treatment, yet its occlusion rate remains suboptimal compared to thermal tumescent techniques. An innovative three-balloons catheter has been developed to allow sclerotherapy in empty vein conditions (empty vein ablation technique, EVA). Aim of this investigation was to describe the EVA technical aspects and related ex-vivo effects on vein wall. METHODS: Two samples from jugular veins of an adult sheep were treated by EVA or foam sclerotherapy (FS, Tessari method). Primary outcome was the percentage of circumferential intima treated by EVA or FS; secondary outcomes were intima and media thickness modifications after treatment. RESULTS: Intact circumferential residual intima were 6.07±2.94% and 16.55±0.70% after EVA and FS, respectively (P=0.020). Despite the average intima and media thickness did not differ between treatments, EVA demonstrated a homogenous damage throughout the vein segment, while FS effect was less destructive distally to the injection site, because moving away from the injection site and floating, it has a less contact with internal surface of the vein. CONCLUSIONS: EVA seems to overcome chemical ablation limits as flushing effect and the increases vein wall/sclerosant agent contact effect compared to FS. Ex-vivo encouraging results need in-vivo validation to evaluate other points like deactivation of sclerosing agent by blood protein and the contact time control between SA and the vein wall. If we have further confirmations in vivo we might think we have a potential higher occlusion rate compared to FS, paving the way for future clinical trials.

Clinical, histological, immunohistochemical, and biomolecular analysis of hyaluronic acid in early wound healing of human gingival tissues: A randomized, split-mouth trial.

BACKGROUND: Hyaluronic acid (HA) exerts a fundamental role in tissue repair. In vitro and animal studies demonstrated its ability to enhance wound healing. Nevertheless, in vivo human studies evaluating mechanisms involved in oral soft tissue repair are lacking. The aim of this study was to evaluate the in vivo effect of HA on early wound healing of human gingival (G) tissues. METHODS: In the present randomized, split-mouth, double-blind, clinical trial, G biopsies were obtained in eight patients 24 h post-surgery after HA application (HA group) and compared with those obtained from the same patients without HA application (no treatment; NT group). Clinical response was evaluated through the Early Wound Healing Score (EHS). Microvascular density (MVD), collagen content and cellular proliferation were evaluated through sirius red and Masson trichrome staining, and Ki-67 immunohistochemistry, respectively. To assess collagen turnover, MMP-1, MMP-2, MMP-9, TGF-ß1 protein levels and LOX, MMP1, TIMP1, TGFB1 gene expression were analyzed by western blot and real time polymerase chain reaction. RESULTS: Twenty-four hours after surgery, the EHS was significantly higher in the HA group. MVD, collagen content, and cell proliferation were not affected. LOX mRNA, MMP-1 protein, and TIMP1 gene expression were significantly upregulated in the HA compared to the NT group. CONCLUSIONS: The additional use of 0.8% HA gel does not modify new blood vessel growth in the early phase of gingival wound healing. Concerning the secondary outcomes, HA seems to enhance extracellular matrix remodeling and collagen maturation, which could drive early wound healing of G tissues to improve clinical parameters.