Effects of different concentrations of bromelain and papain enzymes on shear bond strength of composite resin to deep dentin using an etch-and-rinse adhesive system.

BACKGROUND: The dentin substrate can be modified by proteolytic agents, which may affect the bonding strength of adhesive systems to the treated dentin surface. Papain, a cysteine protease enzyme with antibacterial and anti-inflammatory properties, can be used for deproteinization of dentin. An alternative deproteinizing enzyme is bromelain. OBJECTIVES: This study aimed to evaluate the impact of deproteinization on the shear bond strength (SBS) of composite resin to deep dentin using different concentrations of bromelain and papain. MATERIAL AND METHODS: Sixty upper premolars were extracted and randomly divided into 5 groups (n = 12 per group). In all groups, the dentin surface was etched with 37% phosphoric acid. Group 1 did not receive any enzyme treatment, group 2 was treated with a 10% papain solution, group 3 was treated with a 15% papain solution, group 4 was treated with a 6% bromelain solution, and group 5 was treated with a 10% bromelain solution. After applying an etch-and-rinse adhesive system, the specimens were restored with composite resin and the SBS was measured. RESULTS: Statistically significant differences were found between groups 2 and 3 (10% papain and 15% papain, p = 0.004), groups 2 and 4 (10% papain and 6% bromelain, p = 0.017), groups 4 and 5 (6% bromelain and 10% bromelain, p = 0.021), and groups 3 and 5 (15% papain and 10% bromelain, p = 0.005). CONCLUSIONS: Deproteinization with papain and bromelain at different concentrations after acid etching did not affect the SBS of composite resin to deep dentin when using an etch-and-rinse adhesive system. However, the group deproteinized with 15% papain demonstrated a higher SBS than the group deproteinized with 10% papain, and the group deproteinized with 6% bromelain showed a higher SBS compared to the group deproteinized with 10% bromelain.

Bromelain can reduce the negative effects of a subclinical necrotic enteritis in broiler chickens.

This study was conducted to examine the efficacy of a bromelain-based supplementation coded ANR-pf on growth performance and intestinal lesion of broiler chickens under necrotic enteritis (NE) challenge. A total of 540 Ross 308 day-old male chicks were randomly allocated into 6 treatments of 6 replicates. The bromelain formulation was delivered to chickens through gavaging or in drinking water method twice, on d 8 and 13. Nonchallenged groups included 1) without or 2) with the specific bromelain formulation gavaged at 0.8 mL/kg. NE-challenged groups included 3) without the specific bromelain formulation; 4) gavaged with 0.4 mL/kg; 5) gavaged with 0.8 mL/kg and 6) supplemented with 0.8 mL/kg via drinking water. Birds were challenged with Eimeria spp. on d 9 and Clostridium perfringens (NE-18 strain) on d 14 and 15. On d 14 and 19, fresh faecal contents were collected for the determination of oocyst counts. Intestinal lesion scores were determined on d16. Performance and mortality were recorded throughout the entire experiment. Among challenged groups, birds received additive via drinking water had higher weight gain (WG) compared to the remaining groups (P < 0.001) in the grower phase and had lower FCR compared to 0.4 mL/kg inoculated group in the grower and finisher phases (P < 0.001). Bromelain supplementation via drinking water improved the WG of challenged birds, similar to that of the nonchallenged birds (P < 0.001), and lowered FCR compared to other challenged groups (P < 0.001). Nonchallenged birds and birds that received bromelain formulation in drinking water did not have lesions throughout the small intestine whereas challenged birds, either un-supplemented or supplemented with bromelain via inoculation route recorded similar lesion score levels in the jejunum. At d 19, birds received bromelain in drinking water had lower fecal oocyst numbers compared to challenged birds without additive (P < 0.001). In conclusion, bromelain administration via drinking water could ameliorate the negative impacts of NE-infection in broilers by improving performance, lowering the oocyst numbers and lesion scores.

Effect of the use of bromelain associated with bioactive glass-ceramic on dentin/adhesive interface.

OBJECTIVES: To evaluate the effect of bromelain associated with Biosilicate on the bond strength (BS) of a universal adhesive system to sound (SD) and caries-affected dentin (CAD), and on the proteolytic activity. MATERIALS AND METHODS: Cavities were prepared in 360 molars, half submitted to cariogenic challenge. Teeth were separated into groups (n=20): Control-No treatment; CHX-0.12% chlorhexidine; NaOCl-5% sodium hypochlorite; Br5%-5% bromelain; Br10%-10% bromelain; Bio-10% Biosilicate; NaOClBio-NaOCl+Bio; Br5%Bio-Br5%+Bio; Br10%Bio-Br10%+Bio. Following treatments, the adhesive system was applied, and cavities were restored. Samples were sectioned into sticks and stored at 37 °C for 24 h, 6 months, and 1 year. Microtensile BS (2-way ANOVA, Bonferroni's test, α=0.05), fracture patterns (SEM), and adhesive interfaces (TEM) were evaluated. Bacterial collagenase assay and in situ zymography were performed. RESULTS: In CAD, Br10% presented higher BS (p=0.0208) than Br5%Bio. Br5% presented higher BS (p=0.0033) after 6 months than after 24 h; and association of treatments, higher BS (p<0.05) after aging than after 24 h. Mixed fractures were the most prevalent. Association of treatments promoted a more uniform hybrid layer with embedded Bio particles. Experimental groups presented lower (p<0.0001) relative fluorescence units than Control. Bromelain, associated or not with Bio, showed collagenolytic degradation. CONCLUSIONS: Bromelain associated with Biosilicate did not affect the BS to SD. In CAD, Br5%Bio decreased immediate BS but had no long-term influence. This association decreased the proteolytic activity. CLINICAL RELEVANCE: Bromelain and Biosilicate may enhance the longevity of adhesive restorations by inhibiting endogenous proteases.

Protective effect of bromelain on some metabolic enzyme activities in tyloxapol-induced hyperlipidemic rats.

Elevation of one or more plasma lipids, such as phospholipids, cholesterol esters, cholesterol, and triglycerides, is known as hyperlipidemia. In humans and experimental animals, bromelain, the primary active ingredient isolated from pineapple stems, has several positive effects, including anti-tumor growth, anticoagulation, and anti-inflammation. Hence, the purpose of this study was to determine the possible protective impact of bromelain on some metabolic enzymes (paraoxonase-1, glutathione S-transferase, glutathione reductase, sorbitol dehydrogenase [SDH], aldose reductase [AR], butyrylcholinesterase [BChE], and acetylcholinesterase [AChE]), activity in the heart, kidney, and liver of rats with tyloxapol-induced hyperlipidemia. Rats were divided into three groups: control group, HL-control group (tyloxapol 400 mg/kg, i.p. administered group), and HL+bromelain (group receiving bromelain 250 mg/kg/o.d. prior to administration of tyloxapol 400 mg/kg, i.p.). BChE, SDH, and AR enzyme activities were significantly increased in all tissues in HL-control compared to the control, whereas the activity of other studied enzymes was significantly decreased. Bromelain had a regulatory effect on all tissues and enzyme activities. In conclusion, these results prove that bromelain is a new mediator that decreases hyperlipidemia.

Bromelain-loaded nanocomposites decrease inflammatory and cytotoxicity effects of gliadin on Caco-2 cells and peripheral blood mononuclear cells of celiac patients.

Enzyme therapy can be an appropriate treatment option for celiac disease (CeD). Here, we developed Bromelain-Loaded Nanocomposites (BLNCs) to improve the stability and retention of bromelain enzyme activity. After the characterization of BLNCs, the cytotoxicity of BLNCs was determined on the Caco-2 cell line. The effect of BLNCs on gliadin degradation and the production of pro-inflammatory cytokines and anti-inflammatory molecules in peripheral blood mononuclear cells (PBMCs) obtained from celiac patients were assessed. Furthermore, the expression of CXCR3 and CCR5 genes was measured in CaCo-2 cells treated with gliadin, gliadin-digested with BLNCs, and bromelain. Our study demonstrated that the Bromelain entrapment efficiency in these nanoparticles was acceptable, and BLNCs have no toxic effect on cells. SDS-PAGE confirmed the digestion effect of bromelain released from nanocomposites. When Caco-2 cells were treated with gliadin digested by free bromelain and BLNCs, the expression of CXCR3 and CCR5 genes was significantly decreased. PBMCs of celiac patients treated with Bromelain and BLNCs decreased inflammatory cytokines (IL-1ß, IL-6, TNF-α, and IFN-γ) production compared to untreated PBMCs. This treatment also increased IL-10 and CTLA-4 in PBMCs of CeD patients. According to the promising results of this study, we can hope for the therapeutic potential of BLNCs for CeD.

Bromelain and ficin proteolytic effects on gliadin cytotoxicity and expression of genes involved in cell-tight junctions in Caco-2 cells.

Enzyme therapy for celiac disease (CeD), which digests gliadin into non-immunogenic and non-toxic peptides, can be an appropriate treatment option for CeD. Here, we have investigated the effectiveness of bromelain and ficin on gliadin digestion using in vitro, such as SDS-PAGE, HPLC, and circular dichroism (CD). Furthermore, the cytotoxicity of gliadin and 19-mer peptide before and after digestion with these enzymes was evaluated using the MTT assay in the Caco-2 cell line. Finally, we examined the effect of these treatments along with Larazotide Acetate on the expression of genes involved in cell-tight junctions, such as Occludin, Claudin 3, tight junction protein-1, and Zonulin in the Caco-2 cell line. Our study demonstrated bromelain and ficin digestion effects on the commercial and wheat-extracted gliadin by SDS-PAGE, HPLC, and CD. Also, the cytotoxicity results on Caco-2 showed that toxicity of the gliadin and synthetic 19-mer peptide was decreased by adding bromelain and ficin. Furthermore, the proteolytic effects of bromelain and ficin on gliadin indicated the expression of genes involved in cell-tight junctions was improved. This study confirms that bromelain and ficin mixture could be effective in improving the symptoms of CeD.

The in-vitro development of novel enzyme-based chemo-mechanical caries removal agents.

OBJECTIVES: Bromelain is a potent proteolytic enzyme that has a unique functionality makes it valuable for various therapeutic purposes. This study aimed to develop three novel formulations based on bromelain to be used as chemomechanical caries removal agents. METHODS: The novel agents were prepared using different concentrations of bromelain (10-40 wt. %), with and without 0.1-0.3 wt. % chloramine T or 0.5-1.5 wt. % chlorhexidine (CHX). Based on the enzymatic activity test, three formulations were selected; 30 % bromelain (F1), 30 % bromelain-0.1 % chloramine (F2) and 30 % bromelain-1.5 % CHX (F3). The assessments included molecular docking, Fourier-transform infrared spectroscopy (FTIR), viscosity and pH measurements. The efficiency of caries removal was assessed by DIAGNOdent pen, measuring the excavation time and number of applications, followed by a morphological evaluation of the remaining dentine using scanning electron microscopy (SEM). The results were compared to Brix 3000 as a control. RESULTS: The chloramine and chlorhexidine were chemically compatible with bromelain without compromising the enzyme activity. All experimental formulations showed higher viscosity and pH in comparison to Brix 3000. The DIAGNOdent readings were <20 in all groups, and the lowest readings were observed in F2. The excavation time and number of applications were lowest in F2 and F1. Both F2 and F3 produced smooth dentine surfaces with less tissue debris, but more patent dentine tubules were observed in F1 and F2. CONCLUSIONS: The bromelain-contained formulations showed a potential to be used as chemomechanical caries removal agents in vitro. Further laboratory and clinical studies are needed to validate this claim. CLINICAL SIGNIFICANCE: The bromelain from pineapple stem has broad specificity for cleavage the peptide bonds in denatured protein to facilitate their removal. The study proved the efficiency of this enzyme to remove the dental caries chemomechanically when used alone or conjugated with chloramine and/or chlorhexidine to enhance the disinfecting and cleansing properties.

Bromelain: a review of its mechanisms, pharmacological effects and potential applications.

The utilization of plant-derived supplements for disease prevention and treatment has long been recognized because of their remarkable potential. Ananas comosus, commonly known as pineapple, produces a group of enzymes called bromelain, which contains sulfhydryl moieties. Recent studies have shown that bromelain exhibits a wide range of activities, including anti-inflammatory, anti-diabetic, anti-cancer, and anti-rheumatic properties. These properties make bromelain a promising drug candidate for the treatment of various diseases. The anti-inflammatory activity of bromelain has been shown to be useful in treating inflammatory conditions such as osteoarthritis, rheumatoid arthritis, and asthma, whereas the anti-cancer activity of bromelain is via induction of apoptosis, inhibition of angiogenesis, and enhancement of the body's immune response. The anti-diabetic property of bromelain is owing to the improvement in glucose metabolism and reduction in insulin resistance. The therapeutic potential of bromelain has been investigated in numerous preclinical and clinical studies and a number of patents have been granted to date. Various formulations and delivery systems are being developed in order to improve the efficacy and safety of this molecule, including the microencapsulated form to treat oral inflammatory conditions and liposomal formulations to treat cancer. The development of novel drug delivery systems and formulations has further ameliorated the therapeutic potential of bromelain by improving its bioavailability and stability, while reducing the side effects. This review intends to discuss various properties and therapeutic applications of bromelain, along with its possible mechanism of action in treating various diseases. Recent patents and clinical trials concerning bromelain have also been covered.

Bromelain mitigates liver fibrosis via targeting hepatic stellate cells in vitro and in vivo.

Various therapeutic approaches are conducted for regression of liver fibrosis and prevent possible further carcinogenic transformation. This study was aimed to assess the prospective therapeutic potential of bromelain against thioacetamide (TAA)-induced liver fibrosis using in-vitro and in vivo approaches. In vitro study, HSC-T6 cell line was used to evaluate the effect of bromelain on HSC-T6 cell viability and apoptosis. In vivo, Rats were treated by TAA for 6 weeks for induction of hepatic fibrosis followed by post treatment by different doses of bromelain and silymarin for further 4 weeks to assess the regression of hepatic fibrosis. The in-vitro findings indicated that bromelain hindered the proliferation of HSCs in concentration dependent manner compared with the untreated cells. The in vivo study revealed that treatment of TAA fibrotic rats with different doses of bromelain and silymarin induced a significant restoration in liver function biomarkers, attenuation of oxidative stress, upregulation of total antioxidant capacity and thereby decline of fibrotic biomarkers and improving histopathological and immunohistochemical changes. In conclusion, This study indicates that bromelain can regress TAA induced hepatic fibrosis in rats via inhibiting HSCs activation, α-SMA expression and the ECM deposition in hepatic tissue in addition to its antioxidants pathway, these findings prove the promising therapeutic potential of bromelain as a novel therapeutic approach for chronic hepatic fibrotic diseases.

Cytotoxic properties of unfractionated and fractionated bromelain alone or in combination with chemotherapeutic agents in colorectal cancer cells.

BACKGROUND: Colorectal cancer (CRC) is one of the most lethal cancers worldwide. Long-term survival is not achieved in metastatic CRC despite the current multidisciplinary therapies. Bromelain, a compound extracted from the pineapple plant, has multiple functions and anticancer properties. Previously, bromelain has been chromatographically separated into four fractions. Fraction 3 (F3) exhibits the highest proteolytic activity. The anticancer effects of F3 bromelain in CRC cells is unknown. METHODS: In vitro cytotoxicity was verified through a sulforhodamine B assay. Apoptosis in CRC cells induced by unfractionated or F3 bromelain was examined using Annexin V-FITC/PI staining and Western blot analysis. ROS status, autophagy and lysosome formation were determined by specific detection kit. RESULTS: The cytotoxicity of F3 bromelain in CRC cells was found to be comparable to that of unfractionated bromelain. F3 bromelain induces caspase-dependent apoptosis in CRC cells. Treatment with unfractionated or F3 bromelain increased superoxide and oxidative stress levels and autophagy and lysosome formation. ATG5/12 and beclin-1 were upregulated, and the conversion of LC3B-I to LC3B-II was increased significantly by treatment with F3 bromelain. Treated CQ, autophagy inhibitor, with unfractionated or F3 bromelain enhances the cytotoxic effects. Finally, the combination of unfractionated and F3 bromelain with a routine chemotherapeutic agent (5-fluourouracil, irinotecan, or oxaliplatin) resulted in synergistically higher cytotoxic potency in CRC cells. CONCLUSION: Unfractionated and F3 bromelain inhibits CRC cell proliferation in vitro, and the cytotoxic effects of unfractionated bromelain are equivalent to F3 bromelain. F3 bromelain may be a potential and potent drug for clinical use due to its anticancer efficacy and high synergistic cytotoxicity when combined with a routine chemotherapeutic agent for CRC.

Effect of Nebulized BromAc on Rheology of Artificial Sputum: Relevance to Muco-Obstructive Respiratory Diseases.

Respiratory diseases such as cystic fibrosis, COPD, and COVID-19 are difficult to treat owing to viscous secretions in the airways that evade mucocilliary clearance. Earlier studies have shown success with BromAc as a mucolytic agent. Hence, we tested the formulation on two gelatinous airway representative sputa models, to determine whether similar efficacy exist. Sputum lodged in an endotracheal tube was treated to aerosol N-acetylcysteine, bromelain, or their combination (BromAc). After measuring the particle size of aerosolized BromAc, the apparent viscosity was measured using a capillary tube method, whilst the sputum flow was assessed using a 0.5 mL pipette. Further, the concentration of the agents in the sputa after treatment were quantified using chromogenic assays. The interaction index of the different formulations was also determined. Results indicated that the mean particle size of BromAc was suitable for aerosol delivery. Bromelain and N-acetylcysteine affected both the viscosities and pipette flow in the two sputa models. BromAc showed a greater rheological effect on both the sputa models compared to individual agents. Further, a correlation was found between the rheological effects and the concentration of agents in the sputa. The combination index using viscosity measurements showed synergy only with 250 µg/mL bromelain + 20 mg/mL NAC whilst flow speed showed synergy for both combinations of bromelain (125 and 250 µg/mL) with 20 mg/mL NAC. Hence, this study indicates that BromAc may be used as a successful mucolytic for clearing airway congestion caused by thick mucinous immobile secretions.

Effect of Bromelain on Chronic Unpredictable Stress-induced Behavioral, Biochemical, and Monoamine Changes in Wistar Albino Rat Model of Depression.

BACKGROUND: Bromelain is a complex mixture of protease enzyme extract from the fruit or stem of the pineapple plant and it has a history of folk medicine use. It is known to have a wide range of biological actions and it is most commonly used as an anti-inflammatory agent, though scientists have also discovered its potential as an anticancer and antimicrobial agent, it has been reported to have positive effects on the respiratory, digestive, circulatory systems and potentially on the immune system. OBJECTIVE: This study was designed to investigate the antidepressant potential of Bromelain in the chronic unpredictable stress (CUS) model of depression. METHODS: We studied the antioxidant activity, and neuroprotective effect of Bromelain by analyzing the fear and anxiety behavior, antioxidants, and neurotransmitter levels, and also by analyzing the histopathological changes. Adult male Wistar albino rats were divided into 5 groups, Control; Bromelain; CUS; CUS + Bromelain, CUS + fluoxetine. Animals of the CUS group, CUS + Bromelain group, and CUS + Fluoxetine group were exposed to CUS for 30 days. Animals of the Bromelain group and CUS + Bromelain group were treated orally with 40 mg/kg Bromelain throughout the period of CUS whereas, the positive control group was treated with fluoxetine. RESULTS: Results showed a significant decrease in oxidative stress marker (lipid peroxidation), and the stress hormone cortisol, in Bromelain-treated CUS-induced depression. Bromelain treatment in CUS has also resulted in a significant increase in neurotransmitter levels, which indicates the efficacy of Bromelain to counteract the monamine neurotransmitter changes in depression by increasing their synthesis and reducing their metabolism. In addition, the antioxidant activity of Bromelain prevented oxidative stress in depressed rats. Also, hematoxylin and eosin staining of hippocampus sections has revealed that Bromelain treatment has protected the degeneration of nerve cells by chronic unpredictable stress exposure. CONCLUSION: This data provides evidence for the antidepressant-like action of Bromelain by preventing neurobehavioral, biochemical, and monoamine alterations.

Bromelain does not provide significant analgesic and anti-inflammatory benefits over placebo in cats undergoing ovariohysterectomy.

OBJECTIVE: To evaluate the efficacy and safety of bromelain to control pain and inflammation in cats undergoing ovariohysterectomy. ANIMALS: 30 client-owned cats undergoing ovariohysterectomy. PROCEDURES: In a randomized, blinded clinical study, cats were assigned to receive either oral bromelain suspension (40 mg/kg [18 mg/lb]; BG, n = 15) or placebo solution (0.1 mL/kg [0.045 mL/lb]; PG, 15), which were administered 90 minutes before and 12 hours after surgery. The anesthetic protocol included acepromazine, meperidine, propofol, and isoflurane. Pain and sedation were assessed at various time points up to 24 hours post-extubation using the UNESP-Botucatu multidimensional composite pain scale, the Glasgow feline composite measure pain scale, and a descriptive numerical scale. Surgical wound inflammation was measured at the same time points, using a numeric rating scale. Morphine was administered as rescue analgesia. Laboratory data (urea, creatinine, gamma-glutamyl transferase, alkaline phosphatase, the prothrombin time, and the fecal occult blood) were analyzed preoperatively and 24 hours after surgery. RESULTS: Pain/inflammation scores, and analgesic requirements did not differ between groups. Shorter recovery time and lower sedation scores were recorded during the first hour post-extubation in the BG than the PG. Postoperatively, serum creatinine and gamma-glutamyl transferase were lower in the BG compared to PG. Compared to baseline values, all biochemistry variables decreased at 24 hours in the BG. The prothrombin time and fecal occult blood did not differ between groups or over time. CLINICAL RELEVANCE: Bromelain did not provide significant analgesic and anti-inflammatory benefits over placebo in cats undergoing ovariohysterectomy.

Bromelain: a candidate to enhance wound healing after endonasal surgeries.

OBJECTIVE: In the present study, we investigated the topical bromelain's cytotoxic effects on mouse fibroblast NIH/3T3 cells via cell culture study. MATERIALS AND METHODS: In this cell culture study, Dulbecco's Modified Eagle Medium (DMEM) with fetal bovine serum (FBS, 10%) and penicillin/streptomycin (1%) was used as a cell growth medium for NIH/3T3 mouse fibroblast cells. MTT test was performed in 96-well plates seeded with NIH/3T3 cells 5x103/well and under standard cell culture conditions. Bromelain doses of 3.13 to 100 µM were administered to the wells and incubated for 24, 48, and 72 hours in the same cell culture conditions. For Confocal microscopic evaluation, NIH/3T3 cells were plated on cover slips in 6-well plates (105 cells/well) and treated with 100 µM concentration of bromelain for 24 h. Untreated cells were used as controls. RESULTS: MTT results showed that bromelain is not cytotoxic on mouse fibroblast NIH/3T3 cells. All three incubation times of 24, 48, and 72 hours bromelain initiated cell growth. A statistically significant rise in cell growth was detected in the only applied highest dose of 100 µM bromelain for all incubation times except for 24 hours. The nontoxic effect was further investigated by using confocal microscopy by applying the highest bromelain dose of 100 µM to NIH/3T3 mouse fibroblast cells. Confocal micrographs showed that bromelain did not change the morphology of mouse fibroblast cells at the incubation time of 24h. In untreated cells and bromelain-treated cells, the nucleus of NIH/3T3 cells was undamaged and compact, and the cytoskeleton was fusiform and non-fragmented. CONCLUSIONS: Bromelain is not cytotoxic on mouse fibroblast NIH/3T3 cells and enhances cell growth. If clinical trials will confirm this, it is possible that bromelain will be used topically in humans to enhance wound healing, in rhinosinusitis and chronic rhinosinusitis with nasal polyps and endonasal surgeries due to its anti-inflammatory effects.

Diosmin and Bromelain Stimulate Glutathione and Total Thiols Production in Red Blood Cells.

Diosmin and bromelain are bioactive compounds of plant origin with proven beneficial effects on the human cardiovascular system. We found that diosmin and bromelain slightly reduced total carbonyls levels and had no effect on TBARS levels, as well as slightly increased the total non-enzymatic antioxidant capacity in the RBCs at concentrations of 30 and 60 µg/mL. Diosmin and bromelain induced a significant increase in total thiols and glutathione in the RBCs. Examining the rheological properties of RBCs, we found that both compounds slightly reduce the internal viscosity of the RBCs. Using the MSL (maleimide spin label), we revealed that higher concentrations of bromelain led to a significant decrease in the mobility of this spin label attached to cytosolic thiols in the RBCs, as well as attached to hemoglobin at a higher concentration of diosmin, and for both concentrations of bromelain. Both compounds tended to decrease the cell membrane fluidity in the subsurface area, but not in the deeper regions. An increase in the glutathione concentration and the total level of thiol compounds promotes the protection of the RBCs against oxidative stress, suggesting that both compounds have a stabilizing effect on the cell membrane and improve the rheological properties of the RBCs.

ACE2-Inhibitory Effects of Bromelain and Ficin in Colon Cancer Cells.

Background and Objectives: Bromelain and ficin are aqueous extracts from fruits of Ananas comosus and Ficus carcia plants, used widely for medical applications. Angiotensin-converting enzyme 2 (ACE2) is a homolog of ACE, degrading Ang II to angiotensin 1-7 and decreasing the cellular concentration of Ang II. Materials and Methods: In this study, we investigated the ACE2-inhibitory, antiproliferative, and apoptosis-inducing effects of ficin and bromelain on caco-2 cells. Results: We found that bromelain and ficin significantly reduced the viability of human colon cancer cells with IC50 value concentrations of 8.8 and 4.2 mg/mL for bromelain after 24 and 48 h treatments, and 8.8 and 4.2 mg/mL for ficin after 24 and 48 h treatments, respectively. The apoptosis of the caco-2 cell line treated with bromelain was 81.04% and 56.70%, observed after 24 and 48 h. Total apoptotic proportions in caco-2 cells treated with ficin after 24 and 48 h were 83.7% and 73.0%. An amount of 1.6 mg/mL of bromelain and ficin treatments on caco-2 cells after 24 h revealed a higher decrease than that of other concentrations in the expression of ACE2 protein. Conclusions: In conclusion, bromelain and ficin can dose-dependently decrease the expression of ACE2 protein in caco-2 cells.

A Comparison of Topical Agents for Eschar Removal in a Porcine Model: Bromelain-enriched vs Traditional Collagenase Agents.

Surgical excision and grafting of deep partial-thickness (DPT) and full-thickness (FT) burns is a cornerstone of wound care. The use of commercially available topical enzymatic agents has been limited due to slower and less complete eschar removal than surgical excision. Using a porcine model of DPT and FT burns, we compared the eschar removal efficacy of a bromelain-enriched enzymatic agent derived from the stems of pineapple plants and a commercially available collagenase. We created 40 DPT and 40 FT burns on four anesthetized Yorkshire pigs. Eschar removal was initiated 24 hours later. Two pigs each were randomly assigned to collagenase or the bromelain-enriched agent. The bromelain-enriched agent was applied topically once for 4 hours followed by a 2-hour soaking. The collagenase was applied topically daily until complete removal of eschar or for up to 14 days. All bromelain-enriched treated FT burns underwent complete removal of the eschar after a single application while none of the collagenase-treated FT burns underwent complete removal of the eschar even after 14 days of treatment. All bromelain-enriched treated DPT burns had complete eschar removal after the single application. None of the collagenase-treated DPT burns experienced complete removal of eschar after 10 days; by day 14, 35% had complete eschar removal, 30% had >50% eschar removed, and 35% had <50% eschar removed. We conclude that eschar removal is quicker and more complete with the bromelain-enriched compared with collagenase debriding agent.

Bromelain-based endodontic irrigant: preparation, properties, and biocompatibility: An in-vitro study.

The aim of the study was to determine the properties, efficacy and biocompatibility of combining bromelain enzyme, chlorohexidine and EDTA (BCE) to create a novel endodontic irrigant. Fourier transform infrared spectrometry was performed to confirm the stability of the BCE and direct contact inhibition test was performed to determine antibacterial action. Baseline pH and surface tension of irrigants was compared with determine stability. Subcutaneous injection to dorsal skin of rabbits was graded for presence of inflammation, oedema, granulation and fibrosis. BCE caused less overall irritation, less oedematous and was earlier to heal than 2.5% NaOCl. The pH stability of BCE was also superior to 2.5% NaOCl. A one-way ANOVA test was performed for the direct contact inhibition and microleakage test. A significant difference was determined (p ≤ 0.05) between BCE and 2.5% NaOCl for antibacterial action. BCE irrigant is effective in preparing dentinal surfaces for root canal without adverse effects and promising longevity.

Effects of traditional and novel proteolytic agents on tissue dissolution and dentine microhardness.

The study aimed to evaluate the tissue-dissolving ability of papain and bromelain with respect to that of sodium hypochlorite (NaOCl) at the temperatures of 25°C and 60°C. The study also assessed the effects of these proteolytic agents on radicular dentine microhardness. Warming NaOCl, papain and bromelain solutions resulted in significant tissue dissolution at all time intervals (p < 0.001). At 60°C, bromelain showed significantly higher tissue weight loss at every time interval when compared to NaOCl (p < 0.001). All of the three organic tissue dissolvents reduced the microhardness at 1 hr when compared to their respective baseline values. The reduction in microhardness from the baseline reading was statistically significant only in the papain group at 30 min (p = 0.018) and at 60 min (p = 0.03) when compared to the control group. Hence it was concluded that bromelain exerted superior tissue dissolution action, especially when warmed, with minimal effect on dentine microhardness.

An open-label, proof-of-concept study assessing the effects of bromelain-based enzymatic debridement on biofilm and microbial loads in patients with venous leg ulcers and diabetic foot ulcers.

BACKGROUND: Most chronic wounds contain biofilm, and debridement remains the centerpiece of treatment. Enzymatic debridement is an effective tool in removing nonviable tissue, however, there is little evidence supporting its effect on planktonic and biofilm bacteria. OBJECTIVE: This study evaluated the effects of a novel BBD agent on removal of nonviable tissue, biofilm, and microbial loads in patients with chronic ulcers. MATERIALS AND METHODS: Twelve patients with DFU or VLU were treated with up to 8 once-daily applications of BBD and then followed for an additional 2 weeks. Punch biopsy specimens were collected and analyzed for biofilm, and fluorescence imaging was used to measure bacterial load. RESULTS: Ten patients completed treatment, and 7 achieved complete debridement within a median of 2 applications (range, 2-8). By the end of the 2-week follow-up period, the mean ± SD reduction in wound area was 35% ± 38. In all 6 patients who were positive for biofilm at baseline, the biofilm was reduced to single individual or no detected microorganisms by the end of treatment. Red fluorescence for Staphylococcus aureus decreased from a mean of 1.09 cm² ± 0.58 before treatment to 0.39 cm² ± 0.25 after treatment. BBD was safe and well tolerated. CONCLUSION: Preliminary data suggest that BBD is safe and that it can be used to effectively debride DFU and VLU, reduce biofilm and planktonic bacterial load, and promote reduction in wound size.