Serum and diet long-chain omega-3 fatty acid nutritional status in Chinese elite athletes

Omega-3 polyunsaturated fatty acids (omega-3 PUFAs) are essential for improving the health and performance of athletes. The present study aimed to evaluate the nutritional status of omega-3 PUFAs in Chinese elite athletes by both dietary intake analysis and serum biomarker detection. A cross-sectional analysis of data from 54 elite athletes (24 men and 30 women) from Shanghai professional sports teams was conducted. A food frequency questionnaire (FFQ) was employed to analyze dietary intake, and gas chromatography–mass spectrometry (GC–MS/MS) was conducted to measure serum biomarkers of PUFAs. Correlation analysis was performed to investigate the relationships of PUFA biomarkers with diet, inflammation and oxidative stress. The results showed that the median intake of EPA + DHA among athletes was 132 mg/d, which is lower than the minimum value recommended by dietary guidelines (250 mg/d). The average serum EPA + DHA was 4.0 ± 1.1%, and the ratio of omega-6/omega-3 was 7.7 ± 1.7. Most (96.3%) of the athletes were below the targeted value of serum EPA + DHA, which is associated with a reduction in cardiovascular risk. Correlation analysis showed that the serum EPA + DHA was positively correlated with the long-term dietary intake of EPA + DHA and negatively correlated with inflammatory markers. In conclusion, the serum circulating EPA + DHA and omega-6/omega-3 ratio are effective biomarkers reflecting the nutritional status of PUFAs in athletes. Omega-3 PUFAs have a potential effect on inhibiting inflammatory markers. Hence, it is necessary for Chinese athletes to improve their suboptimal nutritional status of PUFAs through dietary intervention.     

The Effect of Ozone Gas on IL-1β and IL-10 Levels of Gingival Crevicular Fluid in Aggressive Periodontitis Patients

ABSTRACT

Although the use of ozone therapy in dentistry has become widespread, the number of controlled clinical trials evaluating its effectiveness in periodontal therapy is limited. The aim of this study was to evaluate the efficacy of ozone treatment, that is used in concert with scaling and root planning (SRP), on clinical periodontal parameters and to analyze its effect on cytokine levels of GCF in aggressive periodontitis patients. Totally, 27 patients with aggressive periodontitis were randomly selected into groups of treatment with either subgingival SRP followed by application of ozone with a periodontal probe (SRP+ozone) or subgingival SRP followed by irrigation with serum irrigation (SRP-control). The following parameters were evaluated at baseline (T0), and 6 weeks (T1): plaque index (PI); gingival index (GI); probing pocket depth (PPD), clinical attachment loss (CAL), GCF volume, GCF Interleukin-1β (IL-1β), and GCF Interleukin-10 (IL-10) cytokine levels. There was a significant difference in terms of clinical periodontal parameters before and after treatment in both groups. When comparing between groups, there was no significant difference between the treatment methods after 6 weeks with respect to the PI, PPD, CAL, and GCF IL-10 levels. In contrast, GI, GCF volume, and GCF IL-1β levels were statistically significantly different between the two groups at the 6th week after treatment. Application of ozone as an adjunctive therapy to SRP was shown to provide a statistically significant improvement in treatment results compared to SRP plus serum irrigation.     

Enrichment of Omega-3 PUFAs from Fur Seal Oil

The solvent crystallization and the urea complexation of the Uruguayan fur seal oil (Arctocephalus australis Zim.), in order to obtain enriched omega-3 PUFAs concentrates were studied. The fractionation at –6°C of fur seal oil or its fatty acids dissolved in ethanol or acetone, is not suitable to obtain a PUFAs concentrate. Ethanol as a solvent and a two steps process (the second step consists of the addition of urea to the obtained NUCF) is the most useful procedure to obtain a concentrate of high PUFAs content using urea complexation of the free fatty acids from marine oils. When the concentrate is obtained from fur seal fatty acids, the total PUFAs content is of 90% with an overall yield of 17%. The recovery efficiency of total PUFAs is 78%. This procedure is very simple and relatively cheap.     

No Effect of Omega-3 Fatty Acid Supplementation on Cognition and Mood in Individuals with Cognitive Impairment and Probable Alzheimer’s Disease: A Randomised Controlled Trial

Findings from epidemiological and observational studies have indicated that diets high in omega-3 polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) may reduce the risk of cognitive decline and Alzheimer’s disease (AD). To determine if increasing intake of DHA and EPA through supplementation is beneficial to cognition and mood in individuals with cognitive impairment no dementia (CIND) or Alzheimer’s disease (AD) a four month, randomised, double-blind, placebo controlled study was conducted. Fifty-seven participants with CIND and nineteen with AD were randomised to receive either omega-3 PUFAs (600 mg EPA and 625 mg DHA per day) or placebo (olive oil) over a four month period. Elevating depleted levels of EPA and DHA through supplementation in individuals with CIND or AD was found to have negligible beneficial effect on their cognition or mood. These findings confirm an overall negligible benefit of omega-3 PUFA supplementation for those with cognitive impairment and dementia. More intervention studies need to be undertaken with longer study durations and larger sample sizes. It may prove fruitful to examine effects of different doses as well as effects in other dementia subtypes.     

Mammalian-Like Inflammatory and Pro-Resolving Oxylipins in Marine Algae

Oxylipins constitute a family of oxidized fatty acids, that are well known as tissue hormones in mammals. They contribute to inflammation and its resolution. The major classes of these lipid mediators are inflammatory prostaglandins (PGs) and leukotrienes (LTs) as well as pro-resolving resolvins (Rvs). Understanding their biosynthetic pathways and modes of action is important for anti-inflammatory interventions. Besides mammals, marine algae also biosynthesize mammalian-like oxylipins and thus offer new opportunities for oxylipin research. They provide prolific sources for these compounds and offer unique opportunities to study alternative biosynthetic pathways to the well-known lipid mediators. Herein, we discuss recent findings on the biosynthesis of oxylipins in mammals and algae including an alternative pathway to prostaglandin E₂, a novel pathway to a precursor of leukotriene B₄, and the production of resolvins in algae. We evaluate the pharmacological potential of the algal metabolites with implications in health and disease.     

Dietary Intake and Food Sources of Total and Individual Polyunsaturated Fatty Acids in the Belgian Population Over 15 Years Old

Advances in our knowledge of the physiological functions of dietary polyunsaturated fatty acids (PUFAs) have led to an increased interest in food sources and the level of dietary intake of these nutrients. Up to now, no representative data was available for the Belgian adult population. This study aimed to describe data on the intake and food sources of total and individual omega-6 and omega-3 PUFA for the Belgian population over 15 years old. PUFA intakes were assessed for 3,043 Belgian adults, based on two non-consecutive 24 h recalls. Usual intakes were calculated using the multiple source method. The results showed that the intake of linoleic acid (LA) is in accordance with the recommendation for almost all Belgian adults. However, the intake of omega-3 PUFA is suboptimal for a large part of the studied population and also the intake of total PUFA should be increased for a part of the population. The main food source of LA and α-linolenic acid (ALA) was the group of fats and oils (60.6 % for LA and 53.1 % for ALA). Fish and fish products were the most important sources of long chain omega-3 PUFA. Age influenced fatty acids intake, with higher intake of omega-3 PUFA in the older age groups. To fill the gap between the intake and recommendation of total PUFA, and in particular omega-3 PUFA, sustainable strategies and efficient consumer communication strategies will be needed.     

Characterization of CS/PVA/GO electrospun nanofiber membrane with astaxanthin

Tissue engineering has been widely used in regenerative medicine and tissue engineering scaffolds have become a new research direction for periodontal regenerative repair. We aim to develop a biological scaffold material that can support host immunity and promote periodontal regeneration. In this paper, chitosan (CS)/polyvinyl alcohol (PVA)/graphene oxide (GO)/astaxanthin (ASTA) nanofibers membranes were prepared by electrospinning. The nanofibers were characterized by scanning electron microscopy, infrared spectroscopy, mechanical testing, antibacterial testing and cytotoxicity testing. The CS/PVA/GO/ASTA nanofiber membrane had favorable micro-morphology, good mechanical properties and no cytotoxicity. This preliminary study demonstrates that the CS/PVA/GO/ASTA nanofiber membrane can be used for in vivo and in vitro experiments related to periodontal regeneration. The related mechanism of periodontal regeneration will be evaluated in future studies.     

Disentangling the Molecular Mechanisms of the Antidepressant Activity of Omega-3 Polyunsaturated Fatty Acid: A Comprehensive Review of the Literature

Major depressive disorders (MDDs) are often associated with a deficiency in long-chain omega-3 polyunsaturated fatty acids (ω-3 PUFAs), as well as signs of low-grade inflammation. Epidemiological and dietary studies suggest that a high intake of fish, the major source of ω-3 PUFAs, is associated with lower rates of MDDs. Meta-analyses of randomized placebo-controlled ω-3 PUFAs intervention-trials suggest that primarily eicosapentaenoic acid (EPA), but not docosahexaenoic acid (DHA), is responsible for the proposed antidepressant effect. In this review, we dissect the current biological knowledge on EPA and DHA and their bioactive lipid metabolites to search for a pharmacological explanation of this, to date, unexplained clinical observation. Through enzymatic conversion by cyclooxygenase (COX), lipoxygenase (ALOX), and cytochrome P-450 monooxygenase (CYP), EPA and DHA are metabolized to major anti-inflammatory and pro-resolving lipid mediators. In addition, both ω-3 PUFAs are precursors for endocannabinoids, with known effects on immunomodulation, neuroinflammation, food intake and mood. Finally, both ω-3 PUFAs are crucial for the structure and organization of membranes and lipid rafts. While most biological effects are shared by these two ω-3 PUFAs, some distinct features could be identified: (1) The preferential CYP monooxygenase pathway for EPA and EPA derived eicosanoids; (2) The high CB2 receptor affinities of EPA-derived EPEA and its epoxy-metabolite 17,18-EEQ-EA, while the DHA-derived endocannabinoids lack such receptor affinities; (3) The competition of EPA but not DHA with arachidonic acid (AA) for particular glycerophospholipids. EPA and AA are preferentially incorporated into phosphatidylinositols, while DHA is mainly incorporated into phosphatidyl-ethanolamine, -serine and -choline. We propose that these distinct features may explain the superior antidepressant activity of EPA rich ω-3 PUFAs and that these are potential novel targets for future antidepressant drugs.     

Comparing Nanohydroxyapatite Graft and Other Bone Grafts in the Repair of Periodontal Infrabony Lesions: A Systematic Review and Meta-Analysis

Objective: To compare the results of periodontal infrabony lesions treated using nanohydroxyapatite (NcHA) graft with other bone grafts (BGs). Methods: Four electronic databases were searched including PubMed (NLM), Embase (Ovid), Medline, and Dentistry and Oral Sciences (EBSCO). The inclusion criteria included randomised controlled clinical trials (RCTs) and controlled clinical trials (CCTs). The clinical results of NcHA were compared with other BGs. For clinical attachment level (CAL) gain, probing pocket depth (PPD) decrease, and gingival recession (REC) change, weighted averages and forest plots were computed. Results: Seven RCTs fulfilled the selection criteria that were included. When NcHA was compared to other BGs, no clinically significant differences were found in terms of each outcome assessed, except the REC change for synthetic BGs as compared to NcHA. Conclusions: The use of an NcHA graft showed equivalent results compared to other types of BGs. To further validate these findings, future studies are required to compare the NcHA and various BGs over longer time periods and in furcation deficiencies.     

The Effects of Omega-3 Supplementation on Serum Oxidative Stress Parameters in Experimental Periodontitis in an Animal Model

Experimental studies suggest that omega-3 may be beneficial at preventing alveolar bone loss (ABL) by modulating host immune response. The aim of this study is to evaluate the effects of omega-3 supplementation on ABL and serum oxidative biomarkers in a ligature-induced periodontitis model. Twenty-four Wistar albino rats are divided into 3 groups: control (C, n = 8); ligature-induced periodontitis (L, n = 8); and omega-3 plus ligature-induced periodontitis (O+L, n = 8). All rats are fed with ad libitum diet, and O+L group is additionally supplemented with omega-3 fish oil by oral gavage for 44 days. Experimental periodontitis is induced by placement of ligatures around the maxillary second molars of rats in L and O+L groups. ABL is measured histomorphometrically and serum 8-hydroxy-2'-deoxyguanosine, total antioxidant capacity, and total oxidant status (TOS) are analyzed. ABL is higher in L and O+L groups than the C group. Omega-3 supplementation is significantly reduced ABL in group O+L, compared to L group. Furthermore, TOS levels are lower in O+L group than L group. It is suggested that omega-3 administration may reduce ABL and serum TOS levels, which supports the antioxidant role of omega-3 on periodontal disease pathogenesis. Practical applications: Adjunctive use of omega-3 supplementation in periodontitis seems beneficial, although biochemical mechanisms underlying the conflicting results have not been completely understood. On the other hand, oxidative stress has been used as an appropriate target for host modulation therapies in periodontal disease. Omega-3 supplementation in ligature-induced periodontitis in an animal model successfully reduces alveolar bone loss by modulating serum total oxidant status, which may provide a novel pathway in periodontal disease pathogenesis.     

Tissue engineering--the gateway to regenerative medicine

Tissue engineering as an emerging biotechnology sector aims at the in vitro regeneration of diseased tissues and promises to profoundly change medical practice, offering the possibility of regenerating tissues and organs instead of just repairing them (regenerative medicine). Improved healing processes and a higher quality of life are the expected results. This article gives an overview of different technologies for regenerative medicine and presents results of our own current applied research and development. A recent project was successfully closed with the development of a natural biomaterial for soft tissue oral defects. The establishment of an in vitro bioreactor system enabled us to simulate the mechanical and biological environment in a healing wound and to investigate the suitability of different implant materials for the oral tissue regeneration. Moreover, focusing the attention on an alternative method for the intervertebral disc (IVD) regeneration, we established a new tissue engineered approach, based on the three-dimensional (3D) culture of autologous human IVD cells into a polyurethane (PU)-fibrin composite. IVD cells were able to proliferate and, thanks to the 3D conditions, to differentiate expressing the typical native tissue markers. The development of an automated platform was the goal of an additional project, to standardize the cell culture technology, increase the bio-safety and reduce the production costs, moving tissue engineering nearer to clinical application.     

A modular engineering strategy for high-level production of protopanaxadiol from ethanol by Saccharomyces cerevisiae

Ethanol is a more reduced substrate than sugars. Here, ¹³C-metabolic flux analysis (MFA) revealed that ethanol catabolism could supply sufficient acetyl-CoA and reducing equivalent for PPD biosynthesis. Then, we described modular engineering strategy to optimize a multigene pathway for protopanaxadiol (PPD) production from ethanol in Saccharomyces cerevisiae. PPD biosynthesis was divided into four modules: mevalonate (MVA) pathway module, triterpene biosynthesis module, sterol biosynthesis module, and acetyl-CoA formation module. Combinatorially overexpressing every gene in MVA pathway and optimizing metabolic balance in triterpene biosynthesis module led to significantly enhanced PPD production (42.34 mg/L/OD600). In sterol biosynthesis module, fine-tuning lanosterol synthase gene (ERG7) expression using TetR–TetO gene regulation system enabled further production improvement (51.26 mg/L/OD600). Furthermore, increasing cytoplasmic acetyl-CoA supply by overexpressing a Salmonella ACS (acetyl-CoA synthetase gene) mutant ACS_seL641P improved PPD production to 66.55 mg/L/OD600. In 5 L bioreactor, PPD production of the best-performing strain WLT-MVA5 reached 8.09 g/L, which has been the highest titer of plant triterpene produced in yeast. © 2018 American Institute of Chemical Engineers AIChE J, 65: 866–874, 2019     

Halloysite clay nanotube in regenerative medicine for tissue and wound healing

The vital necessity of effective treatment at damaged tissue or wound site has resulted in emerging tissue engineering and regenerative medicine. Tissue engineering has been introduced as an alternative approach for common available therapeutic strategies in the terms of restoring deformed tissue structure and its functionality via the developing of new bio-scaffold. Designed three-dimensional (3D) scaffolds, alone or in combination with bioactive agents, should be able to stimulate and accelerate the development of engineered tissues and provide proper mechanical support during in-vivo implantation and later regeneration process. To cover it up, a series of new bio-structures with higher mechanical strength were designed through the combination of halloysite nanotubes (HNTs) into 3D bio-polymeric networks. HNTs clay mineral with its unique rod-like structure and distinctive chemical surface features, exhibits excellent biocompatibility and biosafety for doping into regenerative scaffolds to enhance their mechanical stiffness and biological performance. In this paper, the ongoing procedures of bone/cartilage tissue engineering and wound healing strategies focusing on the designing of 3D-HNTs bio-composites and their multi-cellular interactions in-vitro and in-vivo preclinical studies are reviewed. Furthermore, the challenges and prospects of 3D-HNTs and HNTs-based functional bio-devices for regenerative medicine are also discussed.     

Resolvins,docosatrienes, and neuroprotectins,novel omega-3-derived mediators,and their endogenous aspirin-triggered epimers

The molecular basis for the beneficial impact of essential omega-3 (n−3) FA remains of interest. Recently, we identified novel mediators generated from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) that displayed potent bioactions identified first in resolving inflammatory exudates and in tissues enriched with DHA. The trivial names resolvin (resolution phase interaction products) and docosatrienes were introduced for the bioactive compounds from these novel series since they possess potent anti-inflammatory and immunoregulatory actions. Compounds derived from EPA carrying potent biological actions (i.e., 1–10 nM range) are designated E series and denoted resolvins of the E series (resolvin E1 or RvE1), and those biosynthesized from the precursor DHA are denoted resolvins of the D series (resolvin D1 or RvD1). The number 1 designates the bioactive compounds in this family (e.g., 1–4). Bioactive members from DHA-containing conjugated triene structures or docosatrienes (DT) that possess immunoregulatory and neuroprotective actions were termed neuroprotectins. Aspirin treatment initiates a related epimeric series by triggering endogenous formation of the 17R-D series resolvins and docosatrienes. These epimers are denoted as aspirin-triggered (AT)-RvD and DT, and possess potent anti-in-flammatory actions in vivo essentially equivalent to their 17S series pathway products. These include five distinct series: (i) 18R resolvins from EPA (i.e., RvE1); (ii) 17R series (AT) resolvins from DHA (RvD1 through RvD4); (iii) 17S series resolvins from DHA (RvD1 through RvD4), (iv) DT from DHA; and (v) their AT form 17R series DT. In this article, we provide an overview of the formation and actions of these newly uncovered pathways and products.     

Evaluation of the physical and mechanical properties of high drug load formulations: Wet granulation vs. novel foam granulation

The purpose of this study was to evaluate the influences of intrinsic drug mechanical properties and different granulation binder delivery processes on the physical and mechanical properties of high drug load granulations after wet granulation. Formulations (80% w/w) of acetaminophen (APAP), metformin and aspirin, which are brittle, viscoelastic, and ductile, respectively; were granulated by high-shear wet granulation. Two modes of binder delivery for wet granulation, either conventional or binder foam, were investigated. Particle size, surface area and pore size of the granulations were characterized. Compacts were prepared at a solid fraction of 0.9 under tri-axial decompression and Hiestand indices (worst-case bonding index (BI_w) and brittle fracture index (BFI)) of the compacts were determined. APAP formulations exhibited the smallest geometric mean particle sizes (d_g) and showed only slight differences in d_g values between the two granulation processes. Binder delivery mode affected mechanical properties of the granulated model drugs differently. Foam granulation appeared to enhance the granule plasticity for APAP while aspirin showed a mixed deformation mechanism based on both its high BI_w and high BFI values. The higher BI_w value for aspirin after foam granulation may be attributed to improved binder distribution among particles during granulation. On the other hand, conventional wet granulation improved the plasticity of metformin as measured by the higher BI_w and lower BFI values. Therefore, conventional wet granulation process conferred advantages in manufacturability and product quality for metformin; as compared to foam granulation which did not enhance plasticity for metformin. Based on this study, a wet granulation process can be selected based on knowledge of the intrinsic drug mechanical properties.     

Coronary heart disease: How do the benefits of ω-3 fatty acids compare with those of aspirin, alcohol/red wine, and statin drugs?

At the present time the primary cause of death of most Americans is cardiovascular disease. Approximately 20 million Americans are currently being treated with some form of statin drugs as a means to lower their blood cholesterol levels, and many of these same people also consume some combination of omega-3 FA, aspirin, and alcohol/red wine because of clinical data indicating that each of these, taken alone, seems to improve mortality. Recent studies with omega-3 FA have demonstrated a positive impact on mortality from coronary heart disease as well as from “all causes,” and this article compares their metabolic benefits with those of aspirin, alcohol/red wine, and statin drugs. The article suggests that these four compounds may have synergistic qualities and that clinical trials to study this possibility are warranted.     

Accelerated Wound Closure of Deep Partial Thickness Burns with Acellular Fish Skin Graft

Thermal injuries are caused by exposure to a variety of sources, and split thickness skin grafts are the gold standard treatment for severe burns; however, they may be impossible when there is no donor skin available. Large total body surface area burns leave patients with limited donor site availability and create a need for treatments capable of achieving early and complete coverage that can also retain normal skin function. In this preclinical trial, two cellular and tissue based products (CTPs) are evaluated on twenty-four 5 × 5 deep partial thickness (DPT) burn wounds. Using appropriate pain control methods, DPT burn wounds were created on six anesthetized Yorkshire pigs. Wounds were excised one day post-burn and the bleeding wound beds were subsequently treated with omega-3-rich acellular fish skin graft (FSG) or fetal bovine dermis (FBD). FSG was reapplied after 7 days and wounds healed via secondary intentions. Digital images, non-invasive measurements, and punch biopsies were acquired during rechecks performed on days 7, 14, 21, 28, 45, and 60. Multiple qualitative measurements were also employed, including re-epithelialization, contraction rates, hydration, laser speckle, and trans-epidermal water loss (TEWL). Each treatment produced granulated tissue (GT) that would be receptive to skin grafts, if desired; however, the FSG induced GT 7 days earlier. FSG treatment resulted in faster re-epithelialization and reduced wound size at day 14 compared to FBD (50.2% vs. 23.5% and 93.1% vs. 106.7%, p < 0.005, respectively). No differences in TEWL measurements were observed. The FSG integrated into the wound bed quicker as evidenced by lower hydration values at day 21 (309.7 vs. 2500.4 µS, p < 0.05) and higher blood flow at day 14 (4.9 vs. 3.1 fold change increase over normal skin, p < 0.005). Here we show that FSG integrated faster without increased contraction, resulting in quicker wound closure without skin graft application which suggests FSG improved burn wound healing over FBD.     

Ozonated Olive Oil Enhances the Growth of Granulation Tissue in a Mouse Model of Pressure Ulcer

The curative effect of ozonated olive oil was evaluated using mouse models of cut wounds and pressure ulcers (decubitus or bedsores). Although ozonated olive oil did not significantly accelerate or decelerate wound contraction in either model, some histological modifications were observed. Ozonated olive oil induced blood coagulation in the hypodermis and cell infiltration in the dermis 1 day after its application. Moreover, it enhanced the formation of granulation tissue 10 days after application. These results indicate that ozonated olive oil promotes granulation tissue formation and is effective in the healing of wounds and pressure ulcers.     

Activin A Promotes Osteoblastic Differentiation of Human Preosteoblasts through the ALK1-Smad1/5/9 Pathway

Activin A, a member of transforming growth factor-β superfamily, is involved in the regulation of cellular differentiation and promotes tissue healing. Previously, we reported that expression of activin A was upregulated around the damaged periodontal tissue including periodontal ligament (PDL) tissue and alveolar bone, and activin A promoted PDL-related gene expression of human PDL cells (HPDLCs). However, little is known about the biological function of activin A in alveolar bone. Thus, this study analyzed activin A-induced biological functions in preosteoblasts (Saos2 cells). Activin A promoted osteoblastic differentiation of Saos2 cells. Activin receptor-like kinase (ALK) 1, an activin type I receptor, was more strongly expressed in Saos2 cells than in HPDLCs, and knockdown of ALK1 inhibited activin A-induced osteoblastic differentiation of Saos2 cells. Expression of ALK1 was upregulated in alveolar bone around damaged periodontal tissue when compared with a nondamaged site. Furthermore, activin A promoted phosphorylation of Smad1/5/9 during osteoblastic differentiation of Saos2 cells and knockdown of ALK1 inhibited activin A-induced phosphorylation of Smad1/5/9 in Saos2 cells. Collectively, these findings suggest that activin A promotes osteoblastic differentiation of preosteoblasts through the ALK1-Smad1/5/9 pathway and could be used as a therapeutic product for the healing of alveolar bone as well as PDL tissue.