Tick attachment cement reviewing the mysteries of a biological skin plug system

The majority of ticks in the family Ixodidae secrete a substance anchoring their mouthparts to the host skin. This substance is termed cement. It has adhesive properties and seals the lesion during feeding. The particular chemical composition and the curing process of the cement are unclear. This review summarizes the literature, starting with a historical overview, briefly introducing the different hypotheses on the origin of the adhesive and how the tick salivary glands have been identified as its source. Details on the sequence of cement deposition, the curing process and detachment are provided. Other possible functions of the cement, such as protection from the host immune system and antimicrobial properties, are presented. Histochemical and ultrastructural data of the intracellular granules in the salivary gland cells, as well as the secreted cement, suggest that proteins constitute the main material, with biochemical data revealing glycine to be the dominant amino acid. Applied methods and their restrictions are discussed. Tick cement is compared with adhesives of other animals such as barnacles, mussels and sea urchins. Finally, we address the potential of tick cement for the field of biomaterial research and in particular for medical applications in future.(VLID)480005

Histomorphometric analysis of the palatal soft tissue as donor region for retrieval of connective tissue grafts

Objectives The soft tissue of the palate is the most frequently used donor side for connective tissue grafts. Various techniques have been described to harvest the connective tissue in anterior and posterior regions of the palate (Hürzeler 1999, Jung 2008, Zucchelli 2010). The present study assessed the histological composition of the soft tissue of the palate in the premolar and tuberosity region and compared the histological composition of connective tissue grafts harvested by two different techniques. Methods Tissue samples of the palatal soft tissue of 10 fresh human dentate cadaver heads were harvested in the premolar and tuberosity region. After histological processing, a histomorphometric analysis on the ratio between epithelium, connective tissue, fatty/glandular tissue, and vascular tissue was performed. Height and composition of the total palatal tissue and of digitally marked grafts (two different harvesting techniques: split-flap- and de-epithelialization-technique) were assessed in both regions (premolar and tuberosity) in an area close and more distant from the teeth (Figure 1). Results The height measurements of the palatal soft tissue ranged from 2.4 to 6.9mm. The main parameters (ratio of connective tissue and fatty/glandular tissue) presented no significant difference between the various regions (close and distant areas in the premolar and tuberosity region; p>0.145; Table 1). But significant differences were detected for the histological compositions of the connective tissue grafts (Table 2); the tissue gained by de-epithelialization in the tuberosity region contained a significantly higher amount of connective tissue, than the tissue gained by split-flap-technique in the premolar region (73.3 vs. 56.5%; p=0.041; Figure 2). Altogether, both, height measurements and composition of the palatal tissue, presented a high inter-individual variability (e.g., percentage of fatty/glandular tissue ranged from 0.04 to 73.8%; Figure 3). Comparison between genders revealed significantly higher values of connective tissue in the premolar region of males (p=0.045); all other parameters presented no relevant gender differences (p>0.077). Conclusions Thus far, the connective tissue harvested in the tuberosity region, which is most often done by de-epithelialization to prevent injury to the greater palatine artery (Figure 1), was clinically described to be denser and more firm compared to the tissue gained in the premolar region (most often by split-flap-technique) (Zuhr 2014). The present study is, to the best of our knowledge, the first to prove this clinical assumption with a histomorphometric analysis. Topic of future clinical trials will be to assess, whether the outcome of root coverage procedures is influenced by the histological composition of the graft (more or less fatty)

Relative Composition of Fibrous Connective and Fatty/Glandular Tissue in Connective Tissue Grafts Depends on the Harvesting Technique but not the Donor Site of the Hard Palate

BACKGROUND: Whether the composition of palatal connective tissue grafts (CTGs) varies depending on donor site or harvesting technique in terms of relative amounts of fibrous connective tissue (CT) and fatty/glandular tissue (FGT) is currently unknown and is histologically assessed in the present study. METHODS: In 10 fresh human cadavers, tissue samples were harvested in the anterior and posterior palate and in areas close to (marginal) and distant from (apical) the mucosal margin. Mucosal thickness, lamina propria thickness (defined as the extent of subepithelial portion of the biopsy containing ≤25% or ≤50% FGT), and proportions of CT and FGT were semi-automatically estimated for the entire mucosa and for CTGs virtually harvested by split-flap (SF) preparation minimum 1 mm deep or after deepithelialization (DE). RESULTS: Palatal mucosal thickness, ranging from 2.35 to 6.89 mm, and histologic composition showed high interindividual variability. Lamina propria thickness (P >0.21) and proportions of CT (P = 0.48) and FGT (P = 0.15) did not differ significantly among the donor sites (anterior, posterior, marginal, apical). However, thicker palatal tissue was associated with higher FGT content (P <0.01) and thinner lamina propria (P ≤0.03). Independent of the donor site, DE-harvested CTG contained a significantly higher proportion of CT and a lower proportion of FGT than an SF-harvested CTG (P <0.04). CONCLUSION: Despite high interindividual variability in terms of relative tissue composition in the hard palate, DE-harvested CTG contains much larger amounts of CT and much lower amounts of FGT than SF-harvested CTG, irrespective of the harvesting site

Scientific Reports / Behavior of human periodontal ligament cells on dentin surfaces ablated with an ultra-short pulsed laser

This study aimed to evaluate the effects of an ultrashort pulsed laser (USPL) (1064 nm, 20 ps, 100 kHz) with different laser fluences (F, 4, 6, 8 J/cm2) and pulse overlaps (PO, 0, 50%) on human periodontal ligament cells (hPDLs) behavior. Dentin samples were ablated with USPL with different combinations of fluences and pulse overlaps; some samples were ablated with an Er:YAG laser (2940 nm, 150 s, 100 mJ/pulse, 5 J/cm2) and some samples were ground with a carbide bur. Then hPDLs were grown on the samples after different treatments. Dentin morphology and cell adhesion were observed with SEM and gene expressions were measured by RT-PCR. The results showed dentin surfaces ablated with USPL when F=4 J/cm2, PO=0, and F=6 J/cm2, PO=0 were partially intact with obvious ridges and valleys and cells on these surfaces grew mostly along the valleys. USPL ablated surfaces in other groups were entirely ablated and cell cluster formation was observed. The RT-PCR results showed an upregulation of osteocalcin of cells grown on the dentin after some laser treatment. It can be concluded that USPL could improve the attachment and differentiation of hPDLs and thus potentially promote periodontal tissue regeneration.(VLID)462660

Effects of Prolyl Hydroxylase Inhibitor L-mimosine on Dental Pulp in the Presence of Advanced Glycation End Products

INTRODUCTION Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. METHODS In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. RESULTS In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. CONCLUSIONS Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization

A Low Cost Implantation Model in the Rat That Allows a Spatial Assessment of Angiogenesis

There is continual demand for animal models that allow a quantitative assessment of angiogenic properties of biomaterials, therapies, and pharmaceuticals. In its simplest form, this is done by subcutaneous material implantation and subsequent vessel counting which usually omits spatial data. We have refined an implantation model and paired it with a computational analytic routine which outputs not only vessel count but also vessel density, distribution, and vessel penetration depth, that relies on a centric vessel as a reference point. We have successfully validated our model by characterizing the angiogenic potential of a fibrin matrix in conjunction with recombinant human vascular endothelial growth factor (rhVEGF165). The inferior epigastric vascular pedicles of rats were sheathed with silicone tubes, which were subsequently filled with 0.2 ml of fibrin and different doses of rhVEGF165, centrically embedding the vessels. Over 4 weeks, tissue samples were harvested and subsequently immunohistologically stained and computationally analyzed. The model was able to detect variations over the angiogenic potentials of growth factor spiked fibrin matrices. Adding 20 ng of rhVEGF165 resulted in a significant increase in vasculature while 200 ng of rhVEGF165 did not improve vascular growth. Vascularized tissue volume increased during the first week and vascular density increased during the second week. Total vessel count increased significantly and exhibited a peak after 2 weeks which was followed by a resorption of vasculature by week 4. In summary, a simple implantation model to study in vivo vascularization with only a minimal workload attached was enhanced to include morphologic data of the emerging vascular tree

Neurofilament distribution in the superior labrum and the long head of the biceps tendon

Abstract Background The postoperative course after arthroscopic superior labrum anterior to posterior (SLAP) repair using suture anchors is accompanied by a prolonged period of pain, which might be caused by constriction of nerve fibres. The purpose was to histologically investigate the distribution of neurofilament in the superior labrum and the long head of the biceps tendon (LHBT), i.e. the location of type II SLAP lesions. Methods Ten LHBTs including the superior labrum were dissected from fresh human specimen and immunohistochemically stained against neurofilament (NF). All slides were scanned at high resolution and converted into tagged image file format, and regions of interest (ROIs) were defined as follows: ROI I—superior labrum anterior to the LHBT origin, ROI II—mid-portion of the superior labrum at the origin of the LHBT, ROI III—superior labrum posterior to the LHBT origin and ROI IV—the most proximal part of the LHBT before its attachment to the superior labrum. The entire images were automatically segmented according to the defined ROIs and measured using a programmed algorithm specifically created for this purpose. The NF-positive cells were counted, and their total size and the area of other tissue were measured separately for the different ROIs. Results Distribution of NF-positive cells in absolute numbers revealed a clear but insignificantly higher amount in favour of ROI I, representing the superior labrum anterior to the LHBT origin. Setting ROI I at 100%, a significant difference could be seen compared to ROI III, representing the superior labrum posterior to the LHBT origin (ROI I vs. ROI III with a p value < 0.05). Conclusions Summarizing, the density of neurofilament is inhomogeneously distributed throughout the superior labrum with the highest number of neurofilament in the anterior superior labrum. Thus, suture placement in type II SLAP repair could play an important role for the postoperative pain-related outcome

Preparation and characterization of a decellularized cartilage scaffold for ear cartilage reconstruction

Scaffolds are widely used to reconstruct cartilage. Yet, the fabrication of a scaffold with a highly organized microenvironment that closely resembles native cartilage remains a major challenge. Scaffolds derived from acellular extracellular matrices are able to provide such a microenvironment. Currently, no report specifically on decellularization of full thickness ear cartilage has been published. In this study, decellularized ear cartilage scaffolds were prepared and extensively characterized. Cartilage decellularization was optimized to remove cells and cell remnants from elastic cartilage. Following removal of nuclear material, the obtained scaffolds retained their native collagen and elastin contents as well as their architecture and shape. High magnification scanning electron microscopy showed no obvious difference in matrix density after decellularization. However, glycosaminoglycan content was significantly reduced, resulting in a loss of viscoelastic properties. Additionally, in contact with the scaffolds, human bone-marrow-derived mesenchymal stem cells remained viable and are able to differentiate toward the chondrogenic lineage when cultured in vitro. These results, including the ability to decellularize whole human ears, highlight the clinical potential of decellularization as an improved cartilage reconstruction strategy

Electrospun poly(ester-Urethane)- and poly(ester-Urethane-Urea) fleeces as promising tissue engineering scaffolds for adipose-derived stem cells.

An irreversible loss of subcutaneous adipose tissue in patients after tumor removal or deep dermal burns makes soft tissue engineering one of the most important challenges in biomedical research. The ideal scaffold for adipose tissue engineering has yet not been identified though biodegradable polymers gained an increasing interest during the last years. In the present study we synthesized two novel biodegradable polymers, poly(ε-caprolactone-co-urethane-co-urea) (PEUU) and poly[(L-lactide-co-ε-caprolactone)-co-(L-lysine ethyl ester diisocyanate)-block-oligo(ethylene glycol)-urethane] (PEU), containing different types of hydrolytically cleavable bondings. Solutions of the polymers at appropriate concentrations were used to fabricate fleeces by electrospinning. Ultrastructure, tensile properties, and degradation of the produced fleeces were evaluated. Adipose-derived stem cells (ASCs) were seeded on fleeces and morphology, viability, proliferation and differentiation were assessed. The biomaterials show fine micro- and nanostructures composed of fibers with diameters of about 0.5 to 1.3 µm. PEUU fleeces were more elastic, which might be favourable in soft tissue engineering, and degraded significantly slower compared to PEU. ASCs were able to adhere, proliferate and differentiate on both scaffolds. Morphology of the cells was slightly better on PEUU than on PEU showing a more physiological appearance. ASCs differentiated into the adipogenic lineage. Gene analysis of differentiated ASCs showed typical expression of adipogenetic markers such as PPARgamma and FABP4. Based on these results, PEUU and PEU meshes show a promising potential as scaffold materials in adipose tissue engineering