Lumbrokinase regulates endoplasmic reticulum stress to improve neurological deficits in ischemic stroke.

Ischemic stroke is characterized by the loss of cerebral blood flow, which frequently leads to neurological deficits. Tissue plasminogen activator is the only therapeutic agent approved to treat ischemic stroke but increases the risk of intracranial hemorrhage and mortality. The fibrinogen-depleting agent lumbrokinase has been used to improve myocardial perfusion in symptomatic stable angina and to prevent secondary ischemic stroke. Lumbrokinase is highly fibrin-specific and only active in the presence of fibrin. Therefore, lumbrokinase has a low risk of hemorrhage due to excessive fibrinolysis. In this study, we aimed to clarify the neuroprotection of lumbrokinase in mice subjected to permanent middle cerebral artery occlusion. Lumbrokinase significantly attenuated infarct volume and improved neurological dysfunction. Lumbrokinase dramatically decreased the expressions of the endoplasmic reticulum (ER) transmembrane receptor protein inositol-requiring enzyme-1 (IRE1) and its downstream transcription factor, XBP-1, caspase-12, and NF-κB activity, thereby significantly inhibiting apoptosis and autophagy and decreasing the NLRP3 inflammasome. Our evidence indicates that post-stroke treatment with lumbrokinase protects against ischemic stroke, thereby regulating ER stress through the collective inhibitory effect of the IRE1 signaling pathways to decrease apoptosis, autophagy, and inflammatory responses. We suggest that lumbrokinase is potential as an adjuvant treatment for ischemic stroke.

Exploring the Fibrin(ogen)olytic, Anticoagulant, and Antithrombotic Activities of Natural Cysteine Protease (Ficin) with the κ-Carrageenan-Induced Rat Tail Thrombosis Model.

Although fibrinolytic enzymes and thrombolytic agents help in cardiovascular disease treatment, those currently available have several side effects. This warrants the search for safer alternatives. Several natural cysteine protease preparations are used in traditional medicine to improve platelet aggregation and thrombosis-related diseases. Hence, this study aimed to investigate the effect of ficin, a natural cysteine protease, on fibrin(ogen) and blood coagulation. The optimal pH (pH 7) and temperature (37 °C) for proteolytic activity were determined using the azocasein method. Fibrinogen action and fibrinolytic activity were measured both electrophoretically and by the fibrin plate assay. The effect of ficin on blood coagulation was studied by conventional coagulation tests: prothrombin time (PT), activated partial thromboplastin time (aPTT), blood clot lysis assay, and the κ-carrageenan thrombosis model. The Aα, Bß, and γ bands of fibrinogen are readily cleaved by ficin, and we also observed a significant increase in PT and aPTT. Further, the mean length of the infarcted regions in the tails of Sprague-Dawley rats was shorter in rats administered 10 U/mL of ficin than in control rats. These findings suggest that natural cysteine protease, ficin contains novel fibrin and fibrinogenolytic enzymes and can be used for preventing and/or treating thrombosis-associated cardiovascular disorders.

Should patients with non-muscle-invasive bladder cancer discontinue fibrin clot inhibitors during bacille Calmette-Guérin?

OBJECTIVE: To determine the impact of fibrin clot inhibitor (FCI) use on oncological outcomes in a large contemporary cohort of patients with non-muscle-invasive bladder cancer (NMIBC) treated with adequate bacille Calmette-Guérin (BCG). PATIENTS AND METHODS: We performed an Institutional Review Board-approved review of patients with NMIBC treated with adequate intravesical BCG, at our institution between 2000 and 2018. FCI use at the time of BCG therapy was recorded for each patient. Patients were stratified according to use of FCI medication. Recurrence- and progression-free survival were analysed using Kaplan-Meier methods and Cox proportional hazard models. RESULTS: Overall, 226 of 526 patients (43.0%) used a FCI: aspirin (205), clopidogrel (38), warfarin (18) and novel oral anticoagulant (NOAC; seven). The use of FCIs did not adversely affect either recurrence- or progression-free survival (P = 0.385 and P = 0.131, respectively). These results did not change when the impact of aspirin, clopidogrel or warfarin/NOAC use on recurrence and progression was evaluated separately. On multivariate analysis, FCI use was neither associated with tumour recurrence nor progression. CONCLUSION: The use of FCIs was not associated with adverse oncological outcomes in a large contemporary cohort of patients receiving adequate intravesical BCG for NMIBC. Based on these results, FCIs may be safely continued during BCG immunotherapy.

The effect of adipose-derived stem cells on enthesis healing after repair of acute and chronic massive rotator cuff tears in rats.

BACKGROUND: Chronic massive rotator cuff tears heal poorly and often retear. This study investigated the effect of adipose-derived stem cells (ADSCs) and transforming growth factor-ß3 (TGF-ß3) delivered in 1 of 2 hydrogels (fibrin or gelatin methacrylate [GelMA]) on enthesis healing after repair of acute or chronic massive rotator cuff tears in rats. METHODS: Adult male Lewis rats underwent bilateral transection of the supraspinatus and infraspinatus tendons with intramuscular injection of botulinum toxin A (n = 48 rats). After 8 weeks, animals received 1 of 8 interventions (n = 12 shoulders/group): (1) no repair, (2) repair only, or repair augmented with (3) fibrin, (4) GelMA, (5) fibrin + ADSCs, (6) GelMA + ADSCs, (7) fibrin + ADSCs + TGF-ß3, or (8) GelMA + ADSCs + TGF-ß3. An equal number of animals underwent acute tendon transection and immediate application of 1 of 8 interventions. Enthesis healing was evaluated 4 weeks after the repair by microcomputed tomography, histology, and mechanical testing. RESULTS: Increased bone loss and reduced structural properties were seen in chronic compared with acute tears. Bone mineral density of the proximal humerus was higher in repairs of chronic tears augmented with fibrin + ADSCs and GelMA + ADSCs than in unrepaired chronic tears. Similar improvement was not seen in acute tears. No intervention enhanced histologic appearance or structural properties in acute or chronic tears. CONCLUSIONS: Surgical repair augmented with ADSCs may provide more benefit in chronic tears compared with acute tears, although there was no added benefit to supplementing ADSCs with TGF-ß3.

Comparison of the mechanical properties of platelet-rich fibrin and ankaferd blood stopper-loaded platelet-rich fibrin.

BACKGROUND AND AIM: Platelet-rich fibrin (PRF) can be named as a natural fibrin-based biomaterial favorable to increasing vascularization and able to guide epithelial cell migration to its surface. The membrane has a significant positive effect on protecting open wounds and accelerating healing. Similar to PRF Ankaferd Blood Stopper (ABS) also has positive effects on wound healing. The aim of this study was to detect if we can improve known physical properties of PRF combining with ABS. This idea was based on the known mechanism of ABS in forming protein network without damaging any blood cells. Materials and Methods: A total of 25 adult rabbits used for collecting 5-7 ml of blood passively with the help of winged blood collection needle to the test tube. Collected samples were centrifuged at 3000 rpm for 10 min. Two similar samples obtained from each animal and one of the samples was placed in 20% ABS 80% saline solution for 5 min. Mechanical properties of the membrane samples were measured using Universal Testing Machine. Results: There is the statistically significant difference between PRF and ABS added PRF in elongation/mm (dL) and elongation/% at break values. Maximum force (fMax) and modulus values did not show any statistically significant differences. CONCLUSION: ABS loaded PRF causes better physical properties. This combination seems to exhibit superior performance when used as a membrane barrier solely. Advanced studies can be done on biological properties of ABS loaded PRF, especially on tissue healing.

Platelet-rich fibrin: a boon in regenerative endodontics.

Research into regenerative dentistry has contributed momentum to the field of molecular biology. Periapical surgery aims at removing periapical pathology to achieve complete wound healing and regeneration of bone and periodontal tissue. Regenerative endodontic procedures are widely being added to the current armamentarium of pulp therapy procedures. The regenerative potential of platelets has been deliberated. Platelet-rich fibrin (PRF) is a wonderful tissue-engineering product and has recently gained much popularity due its promising results in wound healing bone induction. The features of this product are an attribute of platelets which, after cellular interactions, release growth factors and have shown application in diverse disciplines of dentistry. This paper is intended to shed light onto the various prospects of PRF and to provide clinical insight into regenerative endodontic therapy.

Revascularization Induced Maturogenesis of Non-Vital Immature Permanent Tooth Using Platelet-Rich-Fibrin: A Case Report.

The aim of this report is to describe a novel method of revascularization therapy done in a non-vital, immature permanent tooth using Platelet-rich fibrin (PRF),in a recently developed scaffold material to overcome limitations associated with the traditional method of revascularization using natural blood clot. PRF prepared from autologous blood was placed in the root canal and patient was followed up regularly at one, three, six, nine and 12 months for detailed clinical and radiographic evaluation. At 12 months, radiographic examination revealed root elongation, root end closure, continued thickening of the root dentinal walls, obliteration of root canal space, and normal periradicular anatomy. However, more long term prospective trials and histological studies are highly needed before to testify PRF a panacea for the regenerative endodontic therapy in children.

Revascularization of immature, nonvital permanent tooth using platelet-rich fibrin in children.

The purpose of this paper was to present a new approach wherein revascularization of the immature, nonvital permanent tooth was performed using platelet-rich fibrin (PRF) as a novel scaffold material. This was performed after disinfection of the root canal space using triple antibiotic paste followed by placing a PRF membrane in the root canal. The patient was followed up regularly at three-, six-, nine-, and 12-month intervals for review. After 12 months, clinical examination showed negative response to percussion and palpation tests but positive response to cold and electric pulp tests. Radiographic examination revealed continued thickening of the root dentinal walls, narrowing of root canal space, root lengthening, and closure of the root apex with normal periradicular architecture. However, more clinical research using large samples is necessary to prove it advantageous for regenerative endodontic therapy in children.

Autologous platelet concentrate for post-extraction socket healing: a systematic review.

BACKGROUND: Autologous platelet concentrates are claimed to enhance hard and soft tissue healing due to the considerable amount of growth factors that are released after application in the surgical site. However, their actual efficacy for improving tissue healing and regeneration in oral surgery applications is controversial. Tooth extraction socket healing represents a proper model to study the effect of autologous platelet-enriched preparations due to the concomitant occurrence of different processes of both hard and soft tissue healing. PURPOSE: To evaluate the efficacy of platelet concentrates for alveolar socket healing after tooth extraction, by conducting a systematic review. MATERIALS AND METHODS: Medline, Embase and Cochrane Central Register of Controlled Trials were searched using a combination of specific search terms. The last electronic search was performed on 15 June, 2014. Manual searching of the relevant journals and of the reference lists of reviews and all identified randomised controlled trials was also performed. Randomised controlled trials evaluating the effect of a platelet concentrate on fresh extraction sockets were included. Further inclusion criteria were that at least 10 patients were treated (at least 5 per group) and there was a minimum follow-up duration of 3 months. Primary outcomes were postoperative complications, patient satisfaction and postoperative discomfort. Secondary outcomes were any clinical, radiographic, histological and histomorphometric variables used to assess hard and soft tissue healing. Assessment of the methodological quality of the trials was made. RESULTS were expressed as fixed-effects models using mean differences for continuous outcomes and risk ratios for dichotomous outcomes, with 95% confidence intervals (CI). RESULTS: The initial search yielded 476 articles. After the screening process, six articles met the inclusion criteria (199 teeth in 156 patients). Three studies were considered at high risk of bias, two at medium risk and one at low risk. A large heterogeneity in study characteristics and outcome variables used to assess hard tissue healing was observed. A meta-analyses of two studies reporting histomorphometric evaluation of bone biopsies at 3 months' follow-up showed greater bone formation when platelet concentrates were used, as compared to control cases (P <0.001; mean difference 20.41%, 95% C.I. 13.29%, 27.52%). Beneficial effects of platelet concentrates were generally but not systematically reported in most studies, in particular when considering the effects on soft tissue healing and the patient's reported postoperative symptoms like pain and swelling, although no meta-analysis could be done for such parameters. CONCLUSIONS: Although the results of the meta-analysis of the present review are suggestive for a positive effect of platelet concentrates on bone formation in post-extraction sockets, due to the limited amount and quality of the available evidence, they need to be cautiously interpreted. A standardisation of the experimental design is necessary for a better understanding of the true effects of the use of platelet concentrates for enhancing post-extraction socket healing.

Increased vascularization during early healing after biologic augmentation in repair of chronic rotator cuff tears using autologous leukocyte- and platelet-rich fibrin (L-PRF): a prospective randomized controlled pilot trial.

HYPOTHESIS: We hypothesized that arthroscopic rotator cuff repairs using leukocyte- and platelet-rich fibrin (L-PRF) in a standardized, modified protocol is technically feasible and results in a higher vascularization response and watertight healing rate during early healing. METHODS: Twenty patients with chronic rotator cuff tears were randomly assigned to 2 treatment groups. In the test group (N = 10), L-PRF was added in between the tendon and the bone during arthroscopic rotator cuff repair. The second group served as control (N = 10). They received the same arthroscopic treatment without the use of L-PRF. We used a double-row tension band technique. Clinical examinations including subjective shoulder value, visual analog scale, Constant, and Simple Shoulder Test scores and measurement of the vascularization with power Doppler ultrasonography were made at 6 and 12 weeks. RESULTS: There have been no postoperative complications. At 6 and 12 weeks, there was no significant difference in the clinical scores between the test and the control groups. The mean vascularization index of the surgical tendon-to-bone insertions was always significantly higher in the L-PRF group than in the contralateral healthy shoulders at 6 and 12 weeks (P = .0001). Whereas the L-PRF group showed a higher vascularization compared with the control group at 6 weeks (P = .001), there was no difference after 12 weeks of follow-up (P = .889). Watertight healing was obtained in 89% of the repaired cuffs. DISCUSSION/CONCLUSIONS: Arthroscopic rotator cuff repair with the application of L-PRF is technically feasible and yields higher early vascularization. Increased vascularization may potentially predispose to an increased and earlier cellular response and an increased healing rate.

Tratamiento combinado: pautas, indicaciones y perfil de seguridad / Combined treatment: Regimens, indications and safety profile

Las estatinas son la base actual del tratamiento hipolipemiante, pese a lo cual pueden tener limitaciones de eficacia y de seguridad. En pacientes de alto riesgo que no alcanzan objetivos terapéuticos, en los que no toleran las estatinas o en los que tienen dislipidemia aterógena podemos combinar 2 o más fármacos de distintas clases terapéuticas, incluyendo, además de las estatinas, ezetimiba, secuestradores de ácidos biliares, fibratos, niacina o ácidos grasos omega 3 de prescripción. No disponemos aún, sin embargo, de evidencias en cuanto a la disminución de episodios cardiovasculares con estas combinaciones (AU)

Statins are the current basis of lipid-lowering therapy, despite which may have limitations on efficacy and safety. In high risk patients who do not achieve current lipid goals, in those intolerant to statins or those with atherogenic dyslipidemia, it is possible combine two or more lipid lowering drugs, including statins, ezetimibe, bile acid sequestrants, fibrates, niacin and prescription omega-3 fatty acids. However, for most of these combination therapies pivotal data on clinical outcomes are still lacking

Comparative evaluation of autologous platelet-rich fibrin and platelet-rich plasma in the treatment of mandibular degree II furcation defects: a randomized controlled clinical trial.

BACKGROUND: The treatment of molar furcation defects remains a considerable challenge in clinical practice. The identification of clinical measurements influential to treatment outcomes is critical to optimize the results of surgical periodontal therapy. The present study aimed to explore the clinical and radiographical effectiveness of autologous platelet-rich fibrin (PRF) and autologous platelet-rich plasma (PRP) in the treatment of mandibular degree II furcation defects in subjects with chronic periodontitis. MATERIAL AND METHODS: Seventy-two mandibular degree II furcation defects were treated with either autologous PRF with open flap debridement (OFD; 24 defects) or autologous PRP with OFD (25), or OFD alone (23). Clinical and radiological parameters such as probing depth, relative vertical clinical attachment level and horizontal clinical attachment level along with gingival marginal level were recorded at baseline and 9 mo postoperatively. RESULTS: All clinical and radiographic parameters showed statistically significant improvement at both the test sites (PRF with OFD and PRP with OFD) compared to those with OFD alone. Relative vertical clinical attachment level gain was also greater in PRF (2.87 ± 0.85 mm) and PRP (2.71 ± 1.04 mm) sites as compared to control site (1.37 ± 0.58 mm), and relative horizontal clinical attachment level gain was statistically significantly greater in both PRF and PRP than in the control group. CONCLUSIONS: The use of autologous PRF or PRP were both effective in the treatment of furcation defects with uneventful healing of sites.

Autologous platelet-rich fibrin in the treatment of mandibular degree II furcation defects: a randomized clinical trial.

BACKGROUND: Platelet-rich fibrin (PRF), an intimate assembly of cytokines, glycan chains, and structural glycoproteins enmeshed within a slowly polymerized fibrin network, has the potential to accelerate soft and hard tissue healing. This double-masked randomized study is designed to evaluate the effectiveness of autologous PRF in the treatment of mandibular degree II furcation defects compared with open flap debridement (OFD). METHODS: Using a split-mouth design, 18 patients with 36 mandibular degree II furcation defects were randomly allotted and treated either with autologous PRF and OFD or OFD. Plaque index, sulcus bleeding index, probing depth, relative vertical and horizontal clinical attachment level, gingival marginal level, and radiographic bone defect were recorded at baseline and 9 months postoperatively. Comparison between indices between the test and control groups was performed using the paired t test except for plaque index and sulcus bleeding index data, which used the χ(2) test. RESULTS: All clinical and radiographic parameters showed statistically significant improvement at the sites treated with PRF and OFD compared to those with OFD alone. CONCLUSION: Within the limitation of this study, significant improvement with autologous PRF implies its role as a regenerative material in the treatment of furcation defects.

The coronally advanced flap in combination with platelet-rich fibrin (PRF) and enamel matrix derivative in the treatment of gingival recession: a comparative study.

OBJECTIVE: The main objective of this study was to evaluate the clinical effectiveness of platelet-rich fibrin membrane used in combination with a coronally advanced flap (CAF) and to compare it with the use of an enamel matrix derivative (EMD) in combination with a coronally advanced flap in gingival recession treatment. MATERIAL AND METHODS: 20 split-mouth cases of maxillary anterior teeth or bicuspids presenting with Miller Class I or II gingival recession were treated with a CAF combined with a platelet-rich fibrin membrane (PRF group) or with EMD (EMD group) placed under a CAF. The following parameters were measured at baseline and at 12 months post treatment: gingival recession (GR), apicocoronal width of the keratinized tissue (WKT), and probing depth (PD). RESULTS: Complete rot coverage in the PRF group was 65% (13 out of 20 recessions) and 60% in the EMD group (12 out of 20 recessions). GR was 4.10 ± 1.05 mm in the PRF group and 3.90 ± 1.00 mm in the EMD group at baseline, and 1.05 ± 0.45 mm in the PRF group and 1.15 ± 0.65 mm in the EMD group at 12 months. The difference observed between the tow groups at 12 months was statistically significant. Average root coverage was 70.5% in the EMD group and 72.1% in the PRF group. WKT was 1.30 ± 0.56 mm in the EMD group and 1.45 ± 0.86 mm in the PRF group at baseline, and 1.90 ± 0.81 mm in the EMD group and 1.62 ± 0.28 mm in the PRF group at 12 months. The difference observed between the two groups at 12 months was not statistically significant. Twelve-month changes in PD were not significantly different between the two groups. The pain intensity was statistically different between the two groups. The pain intensity was statistically different between groups for the first 5 days, favoring the PRF group. CONCLUSIONS: The present study did not succeed in demonstrating any clinical advantage of the use of PRF compared to EMD in the coverage of gingival recession with the CAF procedure. The EMD group showed a higher success rate in increasing WKT than did the PRF group.

Trial with Platelet-Rich Fibrin and Bio-Oss used as grafting materials in the treatment of the severe maxillar bone atrophy: clinical and radiological evaluations.

OBJECTIVES: The aim of the present study is to assess the implant osteointegration, as well as the course of bone regeneration and healing processes, thanks to the sinus lift procedure and by using PRF as a filling material, in association with the Bio-Oss. MATERIALS AND METHODS: 23 patients, requiring maxillary sinus lift in order to place implants into posterior maxillary region, were involved in this study. Selected cases, in which the height of the residual bone was superior to 5 mm, were adopted for surgical procedure of "one-stage sinus lift" (implant insertion concurrently occured with sinus lift resulting 6-9 months the healing and integration time). Before inserting the implant, a small quantity of filling material was placed in the cavity. For this purpose the bone fragment, stored in saline solution, was employed mixed with Bio-Oss and PRF, after being ground. RESULTS: All patients reported no pain to percussion, no sign of tissue suffering in the soft peri-implant tissues, the presence of an optimal primary stability of the inserted implants and a significant increase in the peri-implant bone density. CONCLUSIONS: In all cases included in this protocol, the Authors observed a successful implant-prosthetic rehabilitation, according with Albrektsson criteria.

The efficacy of fibrin tissue adhesives in pleurodesis in rats.

In search for a new sclerosing agent for pleurodesis, fibrin tissue adhesive is compared to tetracycline for its efficacy in rats. Twenty-four albino Wistar rats were divided into 3 groups. Groups 1, 2, and 3 were given intrapleural isotonic saline, 35 mg/kg tetracycline, and fibrin tissue adhesive with fibrinogen and thrombin concentrations of 30 mg/mL and 10 U/mL, respectively. Rats were evaluated for macroscopic pleural adhesions and mean values of macroscopic scoring were compared among the groups. Fibrin tissue adhesive- and tetracycline-treated rats had significantly more adhesions compared to the control group, whereas fibrin tissue adhesive was more effective for pleurodesis than tetracycline and no deaths or major side effects were observed in any rat. Thus, fibrin tissue adhesive was found as a more effective sclerosing agent than tetracycline for pleurodesis in rats.

Effect of fibrin bandage fibrinogen concentration on blood loss after grade V liver injury in swine.

OBJECTIVE: To determine the effect of fibrinogen concentration of dry fibrin bandages on blood loss after grade V liver injury. METHODS: Twenty-four pigs were used. Grade V liver injuries were induced and treated with dry fibrin bandages containing 0, 4, 8, or 15 mg fibrinogen/cm2. Animals were monitored for 60 minutes. Blood loss, fluid use, hematological data, and hemostasis were assessed. RESULTS: Post-treatment blood losses (mean and 95% confidence interval [CI]) were 1,560 mL (356-6,844), 372 mL (65-2,134), 225 mL (51-992), and 127 mL (22-732) in the 0-, 4-, 8-, and 15-mg groups, respectively. Only the 15-mg group had results significantly lower than the 0-mg group (p < 0.05). Blood loss was negatively related to fibrinogen concentration (p < 0.05). CONCLUSION: Fibrinogen concentration was inversely related to blood loss after grade V liver injury. The 15-mg formulation was the only one that significantly reduced blood loss.

The use of bone-chip grafts fixed with absorbable bone sealant (Absele).

In this study we have investigated the use of bone-chip grafts which were fixed by Absele. In the study 33 adult guinea-pigs were used. Bone grafts for cervical intervertebral fusions were carried out. At the end of the sixth week the bones were examined histopathologically. Both in the control fusion group and in the experiment group the bone grafts were found to be in good condition, but in the free graft group some bone fragments were found in muscle tissue. There was no evidence of foreign body reaction.

Mediastinal fibrin glue: hemostatic effect and tissue response in calves.

There is continued controversy regarding the effectiveness and potential adverse effects of fibrin glue. Thus, we chose to evaluate it in a model of experimental calf aortic valve replacement that has been previously well established. Concentrated fibrinogen and topical thrombin were sprayed to form a thin layer of fibrin glue over the mediastinal tissues of 20 consecutive calves undergoing aortic valve replacement. Chest tube outputs of these animals were compared with those of the preceding 20 consecutive calves undergoing aortic valve replacement without fibrin glue. All procedures were performed by the same surgeon, and no other technical changes were made between the two series. Total postoperative chest tube output (mean +/- standard error) was 553 +/- 50 mL for the calves treated with fibrin glue and 1,155 +/- 103 mL for the control calves (p less than 0.001). On histological examination of mediastinal tissues from 5 treated calves killed 6 weeks after operation, there was no evidence of inflammation, fibrosis, or residual fibrin. To our knowledge, this is the first controlled laboratory study to show that fibrin glue spray is an effective hemostatic agent and that it produces no long-term tissue reaction.