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Although several technologies have been developed to isolate cells of interest from a heterogenous sample, clogging and impaired cell viability limit such isolation. We have developed the Enrich TROVO system as a novel, nonfluidic technology to sort live cells. The TROVO system combines imaging-based cell selection and photo-crosslinking of (gelatin methacrylate) gelMA-hydrogel to capture cells. After capture, cells are released by enzymatic digestion of the hydrogel and then retrieved for downstream analysis or further cell culturing. The system can capture cells with a recovery rate of 48% while maintaining 90% viability. Moreover, TROVO can enrich rare cells 506-fold with 93% efficiency using single step isolation from a 1:104 cell mixture, and can also capture one target cell from 1 million cells, reaching an enrichment ratio of 9128. In addition, 100% purity and 49% recovery rate can be achieved by a following negative isolation process. Compared to existing technologies, the TROVO system is clog-resistant, highly biocompatible, and can process a wide range of sample sizes. • A fluidics free platform to isolate rare cells from complicated samples. • Touchless, high throughput capture via parallel micro light beams. • Image based phenotypes used as sorting criteria. • Flexible isolation strategies such as positive and negative capture. • CTCs, cell pools, cell-cell interaction events can be visualized and isolated. [ABSTRACT FROM AUTHOR]

Tideglusib has shown great performance in terms of dentin regenerative properties. This study aims to evaluate bonding ability, of demineralized dentin infiltrated with polymeric nanoparticles (NPs) doped with tideglusib (TG) (TG-NPs). Dentin conditioned surfaces were infiltrated with NPs and TG-NPs. Bonded interfaces were created and stored for 24 h and then submitted to mechanical, chemical and thermal challenging. The resin-dentin interface was evaluated through a doubled dye fluorescent technique and a calcium chelator fluorophore under a confocal laser scanning microscopy, and by field emission scanning electron microscopy. Dentin surfaces treated with TG-NPs and load cycled produced higher bond strength than the rest of the groups. Immersion of dentin specimens treated with undoped-NPs in collagenase solution attained the lowest microtensile bond strength (MTBS) values. Both porosity and nanoleakage decreased when dentin was infiltrated with TG-NPs, that revealed strong signals of xylenol orange stain at both hybrid layer and dentinal tubules. The presence of NPs, in general, inducted the presence of mineralized interfaces after mechanical loading and thermocycling. Nanoparticles doped with tideglusib promoted the highest dentin bonding efficacy among groups, as they facilitated the maximum bond strength values with creation of mineral deposits at the hybrid layer and dentinal walls. Tideglusib enabled scarce porosity, nanoleakage and advanced sealing among dentin groups. Doping hydrophilic polymeric NPs with tideglusib, infiltrated in etched dentin represents a reproducible technique to create reparative dentin at the resin-dentin interface, by inducing therapeutic bioactivity. [Display omitted] • Tideglusib/TG promoted the highest dentin bonding efficiency among groups. • TG treated dentin created hermetic tubular resin tags and new minerals favoring sealing. • TG facilitated lower porosity and nanoleakage of the resin dentin interface. [ABSTRACT FROM AUTHOR]

• E. alba used in traditional medicine could be adapted and cultivated in France. • Phytochemistry of the plant was studied through NMR and UHPLC-HRMS/MS analysis. • E. alba shows anti-inflammatory, antioxidant and anti-collagenase activities. • Coumestans and phenolic compounds are mainly involved in the studied activities. Natural products are increasingly studied for pharmaceutical and cosmetic applications. In our continuing search for new natural active ingredients for cosmetic uses, we investigated the phytochemistry and studied the bioactivity of Eclipta alba (L.) cultivated in center-Val de Loire Region (CVLR) in France to develop a new anti-aging active ingredient. This plant is known as the false daisy and is traditionally used in Ayurveda for the treatment of digestive, respiratory, hepatic, and cutaneous disorders and for its activity as a hair growth promoter. To highlight the compounds involved in different bioactivities, a metabolomic study of the plant was undertaken. The extract of the aerial parts, obtained by ultrasound-assisted extraction in ethanol, was tested for antioxidant activity and for its inhibitory effect on various enzymes involved in skin aging. The raw extract was fractionated using liquid/liquid extraction (LLE) and Centrifugal Partition Chromatography (CPC), then fraction activity was evaluated to target the main active compounds. HRMS/MS and NMR were used to identify the compounds contained in the active fractions. Some fractions had showed strong collagenase, cyclooxygenase-2 (COX-2) and xanthine oxidase (XO) inhibition. The two most active fractions contained mostly coumestans (demethylwedelolactone (DWL) and wedelolactone (WL)), and other phenolic compounds present in lower amount, which were identified as carriers of the anti-collagenase, anti-cyclooxygenase 2 and anti-xanthine oxidase activity. Therefore, this study suggested that Eclipta alba grown in CVLR has a phytochemistry in line with the description reported in literature and is highly composed of coumestans, phenolic compounds and saponins. The ethanolic extract has shown anti-inflammatory and antioxidant activity and for the first time an anti-collagenase activity has been demonstrated. These activities have been correlated with the presence of wedelolactone (WL) and demethylwedelolactone (DWL), the two majors' compounds, and other phenolics compounds. Thus, Eclipta alba (L.) cultivated in CVLR could be used as an interesting antioxidant and anti-aging agent for cosmetical applications. [ABSTRACT FROM AUTHOR]

The objective of this systematic review and meta-analysis was to synthesize the available randomized controlled trial data comparing needle fasciotomy and collagenase treatment for single-digit Dupuytren contractures with a minimum of 3-year follow-up and determine whether one treatment is superior regarding contracture correction and functional outcomes. A systematic review and meta-analysis was conducted by searching four databases for randomized controlled trials investigating the single-digit treatment outcomes for Dupuytren contracture comparing collagenase treatment and needle fasciotomy with a minimum of 3-year follow-up. The risk of bias of included studies was assessed using the Cochrane risk-of-bias tool. A meta-analysis was performed using a random effects model in anticipation of unobserved heterogeneity. The primary outcome measure was contracture recurrence. Secondary outcome measures included final fixed flexion contracture (FFC), Quick Disabilities of Arm, Shoulder and Hand (Quick DASH) scores, and Unité Rhumatologique des Affections de la Main (URAM) scores. After screening 264 articles, 4 randomized clinical trials were eligible for final inclusion. One trial had a high risk of bias, and two trials had some concern for bias. The final meta-analysis included 347 patients, 169 who underwent collagenase treatment and 178 who underwent needle fasciotomy. No significant differences were noted between the groups in contracture recurrence, FFC, and URAM scores. The pooled data showed a higher Quick DASH score in the collagenase treatment group compared with the needle fasciotomy group, but the observed difference was less than what would be expected to be clinically relevant. Needle fasciotomy and collagenase treatment have similar outcomes with regards to contracture recurrence, final FFC, Quick DASH scores, and URAM scores for the single-digit treatment for Dupuytren contracture at a minimum of 3-year follow-up. Relevant factors that may be considered during the shared decision-making process for treatment selection include surgeon and patient preferences, costs of treatment, and the disparate complication profiles of these two treatments. Therapeutic II. [ABSTRACT FROM AUTHOR]

Dupuytren disease is a benign, progressive fibroproliferative disorder of the hands. To date, only one pharmacotherapy (clostridial collagenase) has been approved for use in Dupuytren disease. There is a great need for additional nonsurgical methods that can be used to either avoid the risks of invasive treatments or help minimize recurrence rates following treatment. A number of nonsurgical modalities have been discussed in the past and continue to appear in discussions among hand surgeons, despite highly variable and often poor or no long-term clinical data. This article reviews many of the pharmacotherapies discussed in the treatment of Dupuytren disease and novel therapies used in inflammation and fibrosis that offer potential treatment options. [ABSTRACT FROM AUTHOR]

Collagenase and protease productions from Aspergillus heteromorphus URM0269 were optimized by submerged fermentation using soybean flour as substrate. Fermentations were performed according to composite design using the concentrations of substrate and yeast extract as the independent variables. The best condition was scaled up in a stirred tank bioreactor to assess the fermentation kinetics. The highest production of both enzymes occurred at concentrations of 2.0 % substrate and 0.1 % yeast extract. Contrariwise, after scale-up, collagenase activity increased from 33.5 to 148.5 U/mL, while the protease decreased from 16.3 to 11.7 U/mL. A. heteromorphus URM0269 showed a maximum growth rate of 0.09 h−1 and yields of protease and collagenase on biomass, after 65 h of 2.70 and 34.22 U/mgx, respectively. Collagenase acted optimally at 40 °C and pH 6.0 on collagen as a substrate and displayed an allosteric trend, achieving a maximum reaction rate of 132.47 U/mL. Thermodynamic parameters of collagen degradation such as Gibbs free energy (74.16 kJ/mol), enthalpy (11.64 kJ/mol), entropy (−199.63 J/K.mol), and activation energy (14.25 kJ/mol) were determined for optimal temperature. These results demonstrated that soybean flour is a potential agroindustrial residue for collagenase production. Furthermore, the collagenase displayed promising biochemical and thermodynamic features for other biotechnological applications. [ABSTRACT FROM AUTHOR]

Injectable collagenase Clostridium histolyticum has been an effective and well-tolerated nonsurgical treatment option for the management of Dupuytren contracture of the hand. The purpose of this study was to determine the efficacy of collagenase injection and adverse event rate in patients who had undergone previous collagenase treatment. A retrospective chart review was performed on 332 patients treated with collagenase injection for Dupuytren contracture by three fellowship-trained hand surgeons at a single institution from 2009 to 2019. Fifty-nine joints in 45 patients underwent repeat collagenase therapy for recurrent contracture in the same digit. Pretreatment and posttreatment total metacarpophalangeal and proximal interphalangeal joint flexion contractures were recorded, with complete correction defined as <5° residual digital flexion contracture. Postmanipulation skin tears and adverse events were recorded. A comparison was made between average contracture improvement after initial collagenase injection and that after repeat injection. Forty-five patients with an average duration of 30 months (range, 6–73 months) between initial and repeat collagenase therapies were identified. The mean improvement after first collagenase injection was 45° ± 24° (39° for metacarpophalangeal joint and 50° for proximal interphalangeal joint) compared with a mean improvement of 43° ± 23° (41° for metacarpophalangeal joint and 44° for proximal interphalangeal joint) after second injection. Although similar complete correction rates and skin tear rates (32.2 % for initial and 30.5% for repeat) were observed between initial (80%) and repeat injections (73%), the occurrence of adverse events was 3 times higher (3.4% for initial and 10.2% for repeat) in the latter group. Collagenase treatment of Dupuytren contracture yields effective total flexion contracture correction. Repeat collagenase treatment of previously treated digits yields similar deformity correction and complete correction rates but a higher incidence of adverse events. Therapeutic IV. [ABSTRACT FROM AUTHOR]

Microbial enzymes have been considered extensively due to their low cost as well as suitable production and stability. Collagenases are proteolytic biocatalysts that have various applications in medicine, industry, and research. In this study, gelatinolytic bacteria were isolated from the soil contaminated with slaughterhouse waste. The isolate with the highest gelatin degradation activity was chosen and characterized with 16 S ribosomal DNA (rDNA) sequence via polymerase chain reaction (PCR) method. The selected isolate was tested for biochemical characteristics. Thereafter, the collagenase production in an industrial medium containing bone powder was investigated for 30 h with the selected bacterium. The enzyme molecular weight was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method, with the enzyme effect on the bone powder morphologically studied via scanning electron microscopy (SEM) analysis. The quantitative ninhydrin method and the qualitative zymography method were applied to measure the enzyme activity. The results indicated that enzyme production by the selected isolate reached its highest level after 25 h of cultivation. The enzyme with about 60 kDa molecular weight could efficiently degrade the collagen of bone powder. [Display omitted] • B. paramycoides MHB was isolated from soil contaminated with slaughterhouse waste. • This bacterium produces collagenase. • It shows potential for industrial collagenolytic enzyme production. [ABSTRACT FROM AUTHOR]

Various studies have reported the antimicrobial, antifungal, and antioxidant effects of Asphodelus species. These species are used in the treatment of various skin-related issues such as acne, abscesses, psoriasis, burns, alopecia, wound healing, and eczema in folk medicine. The purpose of the present study is to determine the antioxidant, antityrosinase, anticollagenase and cytotoxic effects of Asphodelus aestivus and Asphodelus fistulosus as potential dermocosmetic agent, as well as the phenolic substance profiles of the species by Reversed phase high performance liquid chromatography (RP-HPLC). Antioxidant capacity of the species was determined by total phenolic and flavonoid content determination, cupric ion reducing power (CUPRAC), ferric ion reducing power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assays. Enzyme inhibition studies were conducted spectrophotometrically. The cytotoxic activity was analyzed by the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. As a result, it was determined that the methanolic extracts obtained from the species showed strong antioxidant activity. Based on the CUPRAC and DPPH results, A. aestivus was found to be more effective while according to the FRAP results, A. fistulosus exhibited greater effectiveness. A. fistulosus was discovered to be abundant in phenolic substances, while A. aestivus was found to be richer in flavonoids. It has also been observed that the methanolic extracts have a high inhibitory effect on collagenase and tyrosinase enzymes. It is believed that the species can be used as a potential resource in the treatment of skin aging and hyperpigmentation problems in the dermocosmetic industry. It is thought that the data obtained in the study will lead to further studies. [Display omitted] • The methanolic extracts obtained from A. aestivus and A. fistulosus showed strong antioxidant activity. • Tyrosinase and collagenase enzyme inhibition activities of the extracts are significant. • It was observed that the methanol extract of A. aestivus had a higher inhibitory effect on enzymes. • It was found that the extracts did not affect cell viability even at maximum concentration. • In HPLC studies, it was determined that the species contained p-hydroxy benzoic acid, syringaldehyde, benzoic acid, quercetin, vanillic acid and sinapic acid as phenolic compounds. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a fibrotic interstitial lung disease in which collagen progressively deposits in the supporting framework of the lungs. The pathological collagen creates a recalcitrant barrier in mesenchyme for drug penetration, thus greatly restricting the therapeutical efficacy. On the other hand, this overloaded collagen is gradually exposed to the bloodstream at fibrotic sites because of the vascular hyperpermeability, thus serving as a potential target. Herein, pathological collagen targeting and penetrating liposomes (DP-CC) were constructed to deliver anti-fibrotic dual drugs including pirfenidone (PFD) and dexamethasone (DEX) deep into injured alveoli. The liposomes were co-decorated with collagen binding peptide (CBP) and collagenase (COL). CBP could help vehicle recognize the pathological collagen and target the fibrotic lungs efficiently because of its high affinity to collagen, and COL assisted in breaking through the collagen barrier and delivering vehicle to the center of injured sites. Then, the released dual drugs developed a synergistic anti-fibrotic effect to repair the damaged epithelium and remodel the extracellular matrix (ECM), thus rebuilding the lung architecture. This study provides a promising strategy to deliver drugs deep into pathological collagen accumulated sites for the enhanced treatment of IPF. Pathological collagen targeting and penetrating liposomes (DP-CC) target the fibrotic lungs and break through the collagen barrier, thus delivering drugs deep into injured alveoli for enhanced IPF therapy. [Display omitted] [ABSTRACT FROM AUTHOR]

To compare collagenase injection with surgical fasciectomy in Dupuytren disease (DD) for the prevalence of contracture in treated fingers 5 years after treatment. This was a single-center, comparative cohort study comprising 2 cohorts of patients treated for DD in 1 or more of 3 ulnar fingers with collagenase injection (159 patients) or surgical fasciectomy (59 patients). At 5 years after treatment, 13 collagenase-treated and 8 fasciectomy-treated patients had undergone subsequent treatment on the treated fingers and were considered to have current contracture. Of the remaining patients, 112 collagenase-treated patients (128 hands, 180 fingers) and 46 fasciectomy-treated patients (49 hands, 63 fingers) attended follow-up evaluation performed by 2 independent examiners (participation rate 84% and 93%, respectively). We defined current contracture in a treated finger as an active extension deficit of ≥20° in the metacarpophalangeal (MCP) or proximal interphalangeal (PIP) joint or a total (MCP + PIP) active extension deficit (TAED) of ≥30°. We used linear mixed models to analyze differences between the cohorts over time. In the collagenase cohort, current contracture was present in 45 (25%) MCP and 60 (33%) PIP joints, and in the fasciectomy cohort, current contracture was present in 12 MCP (19%) and 30 PIP (48%) joints; a TAED of ≥30° was present in 79 (44%) of the collagenase-treated and 30 (48%) of the fasciectomy-treated fingers. In MCP and PIP joints with ≥20° pretreatment contracture, complete correction was observed in 82 (56%) MCP and 30 (30%) PIP joints in the collagenase cohort and 23 (70%) MCP and 5 (16%) PIP joints in the fasciectomy cohort. There was no statistically significant difference between the 2 cohorts in the TAED change over time. In patients with DD, collagenase injection and surgical fasciectomy improved finger joint contracture over the pretreatment status but had a high prevalence of joint contracture in the treated fingers 5 years after treatment. Therapeutic IV. [ABSTRACT FROM AUTHOR]

Organic-inorganic hybrid nanomaterials are considered as promising immobilization matrix for enzymes owing to their markedly enhanced stability and reusability. Herein, collagenase was chosen as a model enzyme to synthesize collagenase hybrid nanoflowers (Col-hNFs). Maximum collagenase activity (155.58 μmol min−1 L−1) and encapsulation yield (90 %) were observed in presence of Zn(II) ions at 0.05 mg/mL collagenase, 120 mM zinc chloride and PBS (pH 7.5). Synthesized Col-Zn-hNFs were extensively characterized by scanning electron microscopy (SEM), energy dispersive X-ray (EDX), X-ray diffraction (XRD), Fourier transform infrared (FTIR), circular dichroism (CD), fluorescence spectroscopy, dynamic light scattering (DLS) and zeta potential measurements. SEM images showed flower-like morphology with average size of 5.1 μm and zeta potential of −14.3 mV. Col-Zn-hNFs demonstrated superior relative activity across wide pH and temperature ranges, presence of organic solvents and surfactants as compared to its free form. Moreover, Col-Zn-hNFs exhibited excellent shelf life stability and favorable reusability. Col-Zn-hNFs showed the ability to suppress and eradicate fully developed insulin fibrils in vitro (IC 50 = 2.8 and 6.2 μg/mL, respectively). This indicates a promising inhibitory potential of Col-Zn-hNFs against insulin amyloid fibrillation. The findings suggest that the utilization of Col-Zn-hNFs as a carrier matrix holds immense potential for immobilizing collagenase with improved catalytic properties and biomedical applications. • Organic-inorganic hybrid nanoflowers were used for the successful immobilization of Clostridial collagenase (Col-Zn-hNFs) • Synthesized Col-Zn-hNFs were characterized by state-of-the-art techniques (SEM-EDX, ATR-FTIR, CD, XRD, TGA, DLS) • Col-Zn-hNFs demonstrated excellent flower-like morphology, enhance catalytic activity and stability • Col-Zn-hNFs exhibited promising inhibitory potential against insulin amyloid fibrillation [ABSTRACT FROM AUTHOR]

Over the past decade, collagenase treatment and needle fasciotomy (NF) have gained widespread popularity in the treatment of Dupuytren contracture. This prospective study was designed to compare the results of these treatments in terms of clinical and patient-reported outcomes. A prospective, randomized, controlled trial included patients with a contracture of 20° or more in a single metacarpophalangeal joint. Patients were allocated to treatment with either NF or collagenase Clostridium histolyticum. The primary outcome was a reduction in the metacarpophalangeal joint contracture to less than 5°. Secondary outcomes included recurrence, the presence of Dupuytren cords, and changes in patient-reported outcomes. The participants were examined 5 years after the intervention. The study cohort comprised 156 patients divided into 2 equally sized groups. After 5 years, data were collected from 143 (92 %) of the initially enrolled participants. The mean time for the clinical follow-up was 5.1 years. In the remaining cohort without a second procedure, 51% (23 patients) in the collagenase Clostridium histolyticum group and 47% (27 patients) in the NF group still had extension deficits of less than 5°. Among the participants with a successful initial procedure, the recurrence rate was 56% (36 patients) in the collagenase Clostridium histolyticum group and 45% (30 patients) in the NF group. There were no differences between the 2 treatments in regard to passive joint extension, reduction of contracture, range of motion, or patient-reported outcomes. The 5-year outcomes for NF are similar to those for collagenase in terms of sustained correction, recurrence, presence of Dupuytren cords, and patient-reported outcomes for the treatment of metacarpophalangeal joint contractures. Therapeutic I. [ABSTRACT FROM AUTHOR]

Uterine fibroids are stiff, benign tumors containing excessive, disordered collagens that occur in 70-80% of women before age 50 and cause bleeding and pain. Collagenase Clostridium histolyticum (CCH) is a bacterial enzyme capable of digesting the collagens present in fibroids. By combining CCH with injectable drug delivery systems to enhance effectiveness, a new class of treatments could be developed to reduce the stiffness of fibroids, preventing the need for surgical removal and preserving fertility. In this work, we achieved localization of CCH via physical entrapment by co-injecting a thermoresponsive pNIPAM-based polymeric delivery system called LiquoGel (LQG), which undergoes a sol-gel transition upon heating. Toxicity study results for LQG injected subcutaneously into mice demonstrate that LQG does not induce lesions or other adverse effects. We then used rheology to quantify the effects of localized CCH injections on the modulus and viscoelasticity of uterine fibroids, which exhibit gel-like behavior, through ex vivo and in vivo digestion studies. Ex vivo CCH injections reduce the tissue modulus by over two orders of magnitude and co-injection of LQG enhances this effect. Rheological results from an in vivo digestion study in mice show a significant reduction in tissue modulus and increase in tissue viscoelasticity 7 days after a single injection of LQG+CCH. Parallel histological staining validates that the observed rheological changes correspond to an increase in collagen lysis after treatment by LQG+CCH. These results show promise for development of injectable and localized enzymatic therapies for uterine fibroids and other dense tumors. Uterine fibroids are stiff, benign tumors containing high collagen levels that cause bleeding and pain in women. Fertility-preserving and minimally-invasive treatments to soften fibroids are needed as an alternative to surgical removal via hysterectomy. We demonstrate through ex vivo and in vivo studies that co-injecting a thermoresponsive polymer delivery system (LQG) alongside a bacterial collagenase (CCH) enzyme significantly increases treatment effectiveness at softening fibroids through CCH localization. We use rheology to measure the modulus and viscoelasticity of fibroids and histology to show that fibroid softening corresponds to a decrease in collagen after treatment with LQG+CCH. These results highlight the utility of rheology at quantifying tissue properties and present a promising injectable therapy for fibroids and other dense tumors. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • Cotinus coggygria is one of the plant species in terms of cosmeceutical effect. • Active plant species usually have records against skin disorders. • Use of active extracts with different antioxidant mechanisms will be more effective. Plants have been a very rich resource for the development of potentially new drug candidates and also the development of cosmetic products for skincare. The skin produces free radicals or reactive oxygen species through repeated exposure to the sun and these molecules cause oxidative stress and inflammatory reactions in the dermal and epidermal layers of the skin. Matrix metalloproteinases are triggered by aging and in inflamed skin, such as photo-aged skin, skin damage, and wrinkles occurred by collapsing dermal proteins such as collagen and elastin. In the development of plant-based skin whitening products, the first target is to investigate the inhibitors of the tyrosinase enzyme, which provides melanin production. The ethanol extracts of 92 herbal extracts and propolis were tested for their elastase, collagenase, and tyrosinase inhibitory effects using ELISA microtiter assays. Their antioxidant effect was determined using DPPH, DMPD, and NO radical scavenging activity, metal-chelation capacity, FRAP, PRAP, and CUPRAC assays. Total phenolic and flavonoid contents of the extracts were also determined spectrophotometrically. Our results from the screening studies showed that the ethanol extracts prepared from the aerial parts of Lamium purpureum subsp. purpureum, the pericarpium of Garcinia mangostana and Pistacia vera , the flowers of Punica granatum, and the seeds of Vitis vinifera were effective against at least two of these enzymes and therefore, the plant species were selected for further studies. [ABSTRACT FROM AUTHOR]

Dupuytren disease is a fibroproliferative disorder that affects the palmar fascia of the hand and results in varying degrees of nodule and cord formation. Over time, patients may develop progressive contractures, impairing their ability to type, to perform with fine instruments, or to participate in social activities such as shaking hands. Treatment options for Dupuytren contractures include needle aponeurotomy (NA), injection of collagenase Clostridium histolyticum (CCH) with manipulation of the digits, and surgical fasciectomy. Over the past decade, the use of CCH has increased. Recent studies have provided additional data regarding the pathophysiology, indications, outcomes, and costs associated with the treatment for Dupuytren contractures, and this review highlights these advances. [ABSTRACT FROM AUTHOR]

The galactomannan-based gel from Cassia grandis seeds was used to incorporate Penicillium sp. UCP 1286 and commercial collagenases. Experiments were carried out according to a 23-full factorial design to identify the most significant parameters for the incorporation process. The pH of the incorporation solution (pH i), stirring time (t), and initial protein concentration in the crude extract (PC i) were selected as the three independent variables, and the efficiency of collagenase incorporation (E) and collagenolytic activity (CA) after 360 min as the responses. pH i and PC i showed positive statistically significant effects on E , while CA was positively influenced by pH i and t , but negatively by PC i. The fungi collagenase was released from the gel following a pseudo-Fickian behavior. Additionally, no <76 % of collagenase was efficiently incorporated into the gel retaining a high CA (32.5–69.8 U/mL). The obtained results for the commercial collagenase (E = 93.88 %, CA = 65.8 U/mL, and n = 0.10) demonstrated a pseudo-Fickian behavior similar to the fungi-collagenase. The results confirm the biotechnological potential of the gel as an efficient matrix for the incorporation of catalytic compounds; additionally, the incorporation of collagenases was achieved by retaining the proteases CA and releasing them in a controlled manner. [Display omitted] [ABSTRACT FROM AUTHOR]

Our goal was to identify potential anti-aging and anti-hypertensive peptides from the most abundant protein in cheese whey (CW), β-lactoglobulin (BLG), using a multidisciplinary approach. These peptides were generated by three different proteases: trypsin, chymotrypsin and subtilisin. Our bioinformatics approach indicated that trypsin or subtilisin are good candidates for generating anti-hypertensive peptides, whereas chymotrypsin has the highest potential to generate anti-aging fragments, especially anti-elastase. This is mainly due to the presence of chymotryptic peptides with highly similar structures to elastase inhibitors. In addition, we aimed to perform the enzymatic hydrolysis of BLG and evaluate in vitro anti-aging and anti-hypertensive potential by assessing the inhibition to collagenase, elastase and angiotensin-converting enzyme (ACE). We found that chymotrypsin was able to generate a <10 kDa fraction of BLG-hydrolysate that reduces elastase activity (∼15%), which corroborates our interpretation of the in silico data. This fraction also showed reduced ACE activity (∼18%), though statistically similar to the intact protein (∼16%). Peptidomic analysis revealed that peptide sequences with predicted anti-elastase activity were present both in BLG and CW chymotrypsin-hydrolyzed samples, though at higher concentrations in the isolated protein, which may support our experimental results. Lastly, CW was hydrolyzed by chymotrypsin and spray-dried to be used as ingredient in a potentially functional milk-based drink. Hydrolysis significantly impacted the characteristics of the dried powder and the drink containing CW hydrolysate attained better evaluation in sensory analysis. Thus, enzymatic hydrolysis with chymotrypsin generated a potentially functional product that improves bioactivity and functionality of CW. • Chymotrypsin cleaves the peptides with more promising interactions with elastase. • Enzymatic hydrolysis improves bioactivity and functionality of cheese whey. • β-Lactoglobulin demonstrated a novel bioactivity encrypted in its sequence. [ABSTRACT FROM AUTHOR]

Burn wounds contain high levels of protease activity due to the need to remodel the damaged extracellular matrix proteins. While necessary, excessive protease activity can lead to improper wound healing and is associated with increased contraction and fibrosis. No studies to date have investigated the expression changes of all the collagenases and elastases in burn wounds. The present study compares gene expression changes and changes in collagenase and elastase activity between burn wound eschar and normal skin in a pediatric population. Deidentified pediatric tissues were used for these experiments. Burn wound tissue was excised as part of normal standard care within a week from injury; normal skin was removed during elective plastic surgery procedures. RNA-sequencing was performed and significant results were confirmed with qRT-PCR. Activity assays showed a significant increase in both collagenase and elastase activity in the burn wound tissue compared to the normal skin. Western blotting and substrate zymography of tissue homogenates evaluated the results at the protein levels. Four elastases and three collagenases were determined to be significantly upregulated in the wound tissues by both RNA-sequencing and qRT-PCR. Cathepsin V was the only protease that was significantly downregulated. All but one metalloproteinase studied was significantly upregulated. None of the serine proteases were significantly altered in the wound tissues. In conclusion, matrix metalloproteinases appear to be the most highly elevated proteases after a pediatric burn wound injury, at least within the first 3-7 days. The data warrant further investigation into the effects of MMPs on burn wound healing. [ABSTRACT FROM AUTHOR]

Cartilage functionality is determined by tissue structure and composition. If altered, cartilage is predisposed to premature degeneration. This pathomimetical study of early osteoarthritis evaluated the dose-dependant effects of collagenase-induced collagen disintegration and proteoglycan depletion on cartilage functionality as assessed by serial T1, T1ρ, T2, and T2* mapping under loading. 30 human femoral osteochondral samples underwent imaging on a clinical 3.0 T MRI scanner (Achieva, Philips) in the unloaded reference configuration (δ 0) and under pressure-controlled quasi-static indentation loading to 15.1 N (δ 1) and to 28.6 N (δ 2). Imaging was performed before and after exposure to low (LC, 0.5 mg/mL; n = 10) or high concentration (HC, 1.5 mg/mL; n = 10) of collagenase. Untreated samples served as controls (n = 10). Loading responses were determined for the entire sample and the directly loaded (i.e. sub-pistonal) and bilaterally adjacent (i.e. peri‑pistonal) regions, referenced histologically, quantified as relative changes, and analysed using adequate parametric and non-parametric statistical tests. Dose-dependant surface disintegration and tissue loss were reflected by distinctly different pre- and post-exposure response-to-loading patterns. While T1 generally decreased with loading, regardless of collagenase exposure, T1ρ increased significantly after HC exposure (p = 0.008). Loading-induced decreases in T2 were significant after LC exposure (p = 0.006), while changes in T2* were ambiguous. In conclusion, aberrant loading-induced changes in T2 and T1ρ reflect moderate and severe matrix changes, respectively, and indicate the close interrelatedness of matrix changes and functionality in cartilage. Image, graphical abstract [ABSTRACT FROM AUTHOR]

• Four compounds were isolated from the plant with activity-guided fractionation. • All the ligands can be regarded as inhibitor candidates with fair binding energies. • Methyl gallate and astragalin were predicted as non-mutagenic. • Cotinus coggygria would be a suitable raw herbal material for cosmetic products. Cotinus coggygria Scop. (Anacardiaceae, syn: Rhus cotinus L.) is known as "boyacı sumağı, sarı boya, duman ağacı" in Turkish and "smoke tree" in English. It is commonly grown in Southern Europe and Anatolia. The leaves have been used due to its antiseptic, hemostatic, antipyretic, and wound healing effects as a 5% infusion in traditional medicine. It has also been reported to be used against skin disorders in Russia. Based on this information, the ethanol extracts prepared from the pedicels and leaves of C. coggygria were investigated for their elastase, collagenase, and tyrosinase inhibitory effects, which are enzymes related to anti-aging, using ELISA microtiter assays. Based on our results, the ethanol extracts prepared from the leaves and pedicels of C. coggygria had low elastase (28.16% ± 2.91 and 25.76% ± 1.71, respectively), moderate collagenase (47.78% ± 4.90 and 46.51% ± 3.15, respectively), and tyrosinase (57.94% ± 0.67 and 46.20% ± 0.92, respectively) inhibition at final concentration (666 µg/mL). The ethanol extract prepared from the pedicels of C. coggygria was subjected to bioactivity-guided fractionation, which led to isolation of methyl gallate, astragalin, isoquercetin, and hyperoside from the active fractions. In addition to the enzyme assays, in order to understand the inhibition mechanisms of the compounds inside the ligand-binding domains, the interactions were simulated and the key amino acids contributing to the hydrogen bonds and non-polar interactions with the ligands were reported. [ABSTRACT FROM AUTHOR]

Dupuytren's contracture is a common condition that has the potential to be debilitating. It presents in a variety of manners and can be mild or more aggressive in its progression. There are a large number of management options currently available. In this review of the evidence, non-operative and operative management options are examined, with a consideration of post-operative rehabilitation and complications. A summary of the current concepts in the management of Dupuytren's contracture is presented. [ABSTRACT FROM AUTHOR]

Successful clinical human allo or auto-islet transplantation requires the recovery of a sufficient number of functional islets from either brain-dead or chronic pancreatitis pancreases respectively. In the last two decades (2000–2019), significant progress has been made in improving the human islet isolation procedures and in standardizing the use of different tissue dissociation enzyme (TDE; a mixture of collagenase and protease enzymes) blends to recover higher islet yields. This review presents information focusing on properties and role of TDE blends during the islet isolation process, particularly emphasizing on the current developments, associated challenges and future perspectives within the field. • Successful human islet isolation and transplantation indicate importance of intact class 1 collagenase. • Recombinant rC1rC2 collagenases have beneficial effects on human islet yield and function. • Increasing protease to collagenase dose improves recovery of human islets from young donor pancreases. • Low-cost enriched collagenase-purified protease enzyme mixtures for successful human islet isolation. [ABSTRACT FROM AUTHOR]

Tissue homeostasis depends on a balance of synthesis and degradation of constituent proteins, with turnover of a given protein potentially regulated by its use. Extracellular matrix (ECM) is predominantly composed of fibrillar collagens that exhibit tension-sensitive degradation, which we review here at different levels of hierarchy. Past experiments and recent proteomics measurements together suggest that mechanical strain stabilizes collagen against enzymatic degradation at the scale of tissues and fibrils whereas isolated collagen molecules exhibit a biphasic behavior that depends on load magnitude. Within a Michaelis-Menten framework, collagenases at constant concentration effectively exhibit a low activity on substrate fibrils when the fibrils are strained by tension. Mechanisms of such mechanosensitive regulation are surveyed together with relevant interactions of collagen fibrils with cells. • Enzymatic degradation of tissues is dependent on mechanical load. • Tensile strain inhibits enzymatic degradation of ECM's collagen at different structural hierarchy levels of most tissues. • At molecular level, collagen degradation rate shows mechanical load magnitude dependent biphasic behavior. • Tensile load suppresses enzymatic degradation of collagen by MMPs and BC. [ABSTRACT FROM AUTHOR]

Plantago major L. which is a medicinal plant with important biological activities, commonly used as traditional medicine. Potential inhibitory activities of the aqueous extract and three isolated constituents calceorioside B (1), homoplantaginin (hispidulin-7-O-glucoside) (2) and plantamajoside (3) from the aerial parts of Plantago major subsp. major L. (Plantaginaceae) have been tested against hyaluronidase, collagenase, and elastase, which play critical roles in wound pathogenesis. Even though, the extract (27.04%), and among the isolated compounds, calceorioside B (41.16%) exerted significant inhibition against hyaluronidase enzyme, homoplantaginin and plantamajoside were found to be inactive. Similar results were obtained from collagenase enzyme inhibition test. The extract (21.92%) and calceorioside B (28.34%) also caused notable inhibition in this test. However, no remarkable inhibition was observed in the presence of elastase enzyme. The experimental data revealed that P. major subsp. major displayed remarkable inhibitory activity against hyaluronidase and collagenase enzymes. In vitro enzyme activity of P. major subsp. major is reported for the first time in the current study. [ABSTRACT FROM AUTHOR]

Withania somnifera (L.) Dunal is widely used in the Indian traditional system of medicine to promote general health, wellness, and longevity. Its pharmacological properties are attributed to a group of molecules called withanolides, among which Withaferin A holds great interest for its anti-carcinogenic action. For this reason, numerous studies in recent years have focused on different metabolic or genetic engineering solutions to increase its yield. Here, we present the Sardinian chemotype of Withania somnifera as a potential crop for the extraction of Withaferin A. W. somnifera was cultivated from Sardinian wild germplasm collected in the northeast of the island. After 18 months, the leaves and the roots were collected and their methanolic extract was analyzed by HPLC. 0.3 mg/g DW of Withanolide A (WA), 1.0 mg/g DW of Withanolide B (WB) and 17.7 mg/g DW of Withaferin A (WF) were detected in the leaf sample, while lower values were detected in the roots (0.1 mg/g WF, 0.3 WA mg/g, 0.1 mg/g WB, 0.2 mg/g WO). This research not only confirms the high Withaferin A content found in the wild population leaves, but shows how they are reproducible in cultivated specimens, highlighting Sardinian W. somnifera leaves as a potential source of high-content Withaferin A products. Finally, we focused on the leaves extract by characterizing the phenolic and flavonoid content, as well as the in-vitro antioxidant capacity by DPPH and ABTS assays, revealing a significant amount of phenolic compounds and a related free radical scavenging activity. The leaves extract was further characterized for its anti-aging properties for potential cosmetic application, by the inhibition of tyrosinase, elastase, and collagenase enzymes. [Display omitted] • Sardinian Withania somnifera was cultivated in open-field. • Leaves extract shows a high content of Withaferin A. • Sardinian W. somnifera represents a potential crop source for Withaferin A production. [ABSTRACT FROM AUTHOR]

• Intramuscular connective tissue (IMCT) was mainly degraded in first 10 days. • IMCT degradation was mainly attributed to decorin. • IMCT was hydrolyzed by endogenous collagenase rather than cathepsins. • IMCT and myofibrils (MF) contributed equally to fish texture maintenance. • IMCT failed to protect MF in the process of texture degradation. Although intramuscular connective tissue (IMCT) is low in fish, its impact on texture cannot be ignored due to its special location. Therefore, this study was aimed to investigate the contribution of IMCT degradation to fish softening and its mechanism induced by endogenous proteases. Results showed that IMCT honeycomb-like structure collapsed entirely on the 10th day of ice storage, along with a decrease of shear force by 36.5%. Meanwhile, IMCT and myofibrils (MF) degradation accelerated softening by 25.1% and 15.3% during 10 days of ice storage, respectively. Next, IMCT deterioration was indicated to be highly correlated with decorin degradation (0.956**), followed by elastin (0.928**) and collagen (0.904**). Ulteriorly, endogenous collagenase was shown to degrade IMCT crucial components, while endogenous cathepsins had little effect. In conclusion, this study confirmed that IMCT played an essential role in maintaining fish texture and was mainly degraded by endogenous collagenase. [ABSTRACT FROM AUTHOR]

Mechanical strain inhibits bacterial collagenase from cleaving collagen. Additionally, the toe region of a soft tissue's force–elongation curve arises from sequentially engaging collagen fibrils as the tissue lengthens. Together, these phenomena suggest that mechanical strain may gradually inhibit collagenase activity through a soft tissue's toe region. Therefore, this investigation sought to test this hypothesis. 92 rat tail tendon fascicles from 3 female sentinel animals underwent preliminary stiffness tests, and their force–elongation curves were fit to a collagen distribution model. This distribution-based model calculated the force magnitude corresponding to p % of collagen fibril engagement. Specimens were separated into one of five levels of p , and that level of force was maintained for two hours while being exposed to 0.054 U/mL of bacterial collagenase from C. histolyticum. The specimens were strained to failure following the creep test, and the relative reduction in stiffness was quantified to estimate the fraction of digested fibrils. Every 10% additional collagen engagement corresponded to a 6.3% (97% highest density interval: 4.3 – 8.4%) retention of stiffness, which indicated collagenase inhibition. The results of this investigation were consistent with a strain-inhibition hypothesis along with the established uncrimping mechanism in the toe region. These results support an interaction between mechanical strain and collagenolysis, which may be valuable for disease prevention or treatment. [ABSTRACT FROM AUTHOR]

The present study aims to identify specific staining methods for acoustic histology. We compared attenuation-of-sound (AOS) images from scanning acoustic microscopy (SAM) with light microscopy (LM) images. Ethanol-fixed tissue or cytology samples and formalin-fixed surgical or autopsy specimens were examined. Nuclei, collagen, elastic fibers and polysaccharides and various antigens, including cell surface, cytoplasmic, nuclear and stromal substances, were observed. Samples with various fixation methods were used. Hematoxylin staining had significantly higher AOS values in accordance with staining duration. Specific staining for collagen, elastic fibers and polysaccharides increased the AOS values of the specific substance. Using diaminobenzidine tetrahydrochloride in NiCl2 solution as a substrate for horseradish peroxidase increased the AOS values to those suitable for acoustic immunostaining. Collagenase digestion after collagen staining decreased AOS values, reflecting collagen density and distribution. Staining with specific dyes or acoustic immunostaining enabled the histologic localization of specific substances by SAM, similar to LM. [ABSTRACT FROM AUTHOR]

Scanning acoustic microscopy reveals information on histology and speed of sound (SOS) through tissues. Slower SOS corresponds to lower stiffness. The aim of the present study was to investigate whether SOS values reflect the degree of degeneration with aging or dissection and whether enzymatic digestion susceptibility is distinct. The SOSs of media other than the atheromatous areas of normal and surgical dissections were measured and compared using medial degeneration grade (MDG) scores. To evaluate the damage rate, SOS was assessed after collagenase digestion. SOS scores negatively correlated with aging and MDG scores. Dissected aortas had higher SOS and MDG scores without age correlation. Collagenase digestion was present in all aortas, but older aortas were more injured than younger aortas. Dissected aortas were more vulnerable to collagenase. Older and dissected aortas expressed specific extracellular matrix components to compensate for mechanical weakness. The present method can evaluate mechanical weakness corresponding to histology to investigate the cause of rupture. [ABSTRACT FROM AUTHOR]

Despite the enormous advances in the field of clinical pancreatic islet transplantation over the past two decades, the human islet isolation procedure remains suboptimal. Islets are extracted (isolated) from the exocrine tissue of donor pancreases using neutral protease (NP) and collagenase-based enzymes, which digest the extracellular matrix (ECM) scaffold surrounding human islets. This process remains highly variable and current isolation enzyme blends are ineffective at digesting pancreases from younger donors with low body mass indexes (BMI). However, age-related differences in pancreatic matrix digestion have not been studied in detail at the molecular level. To address this, we investigated ECM digestion in purified ECM proteins and in pancreatic tissue sections from younger (≤30 years; n = 5) and older (>55 years; n = 5) BMI matched donors, using Raman microspectroscopy (RMS). The Raman spectral profiles for purified collagens I, IV, VI and laminins were significantly altered following controlled enzyme treatment. Pancreatic cryosections were treated with Serva collagenase, NP, or the two enzymes combined, at clinically relevant concentrations. RMS demonstrated that the ECM at the islet-exocrine interface was differentially digested with respect to donor age. The action of collagenase was affected to a greater extent than NP. RMS is a powerful, marker-independent technology for characterising the human pancreatic ECM and demonstrating differences between donor types. Ongoing detailed studies using RMS will assist the development of donor-specific enzyme blends, increasing the overall success of human islet isolation and benefiting many people with type 1 diabetes worldwide. Pancreatic islet transplantation is a minimally invasive treatment, which can reverse Type 1 Diabetes Mellitus (T1DM) in selected patients. Islets of Langerhans are extracted (isolated) from the exocrine tissue of human donor pancreases using neutral protease (NP) and collagenase-based enzymes, which digest the extracellular matrix (ECM) scaffold surrounding human islets. This process remains highly variable and current enzymes are ineffective at digesting pancreases from younger donors. Using Raman microspectroscopy we demonstrate that donor age affects the enzymatic digestion of the pancreatic ECM at the molecular level. Collagenase activity is affected to a greater extent than NP. These findings will assist the development of donor-specific enzymes, thereby increasing the overall success of islet isolation and benefiting many people with T1DM worldwide. [ABSTRACT FROM AUTHOR]

The aims of our study were to evaluate the rates and predictors of reinterventions and direct costs of 3 common treatments of Dupuytren contractures—needle aponeurotomy, collagenase injection, and surgical fasciectomy. A retrospective review identified 848 interventions for Dupuytren contracture in 350 patients treated by a single surgeon from 2005 to 2016. The treatments included needle aponeurotomy (NA) (n = 444), collagenase injection (n = 272), and open fasciectomy (n = 132). We collected information on demographics, contracture details, and comorbidities. Outcomes included reintervention rates, time to reintervention, and direct cost of treatments. Standardized costs were calculated by applying 2017 Medicare reimbursement to professional services and cost-to-charge ratios to hospital charges. Demographics were similar among the 3 treatment groups. The fifth finger was the most commonly affected digit including 43% of the NA, 60% of the collagenase, and 45% of the fasciectomy groups. The 2-year rates of reintervention following NA, collagenase, and fasciectomy were 24%, 41%, and 4%, respectively, and the 5-year rates were 61%, 55%, and 4%, respectively. Younger age and severity of preintervention proximal interphalangeal (PIP) joint contracture were predictive of reintervention in the NA and collagenase groups. The standardized direct costs for NA, collagenase, and fasciectomy were $624, $4,189, and $5,291, respectively. Including all reinterventions, the cumulative costs per digit following NA, collagenase, and surgery at 5 years were $1,540, $5,952, and $5,507, respectively. Treatment with collagenase resulted in the highest rate of reintervention at 2 years, comparable reintervention rates to NA at 5 years, and the highest cumulative costs. The NA was the least expensive and resulted in longer duration before reintervention compared with collagenase. More severe PIP joint contractures and younger age at time of initial intervention were predictive of reintervention after collagenase and NA. Fasciectomy has a high initial cost but the lowest reintervention rate. Therapeutic IV. [ABSTRACT FROM AUTHOR]

3-methoxysalicylidene N-alkyl-thiosemicarbazones (L) (where alkyl is propyl, butyl, pentyl or hexyl) were synthesized. The reaction of the ONS donor ligands with [MoO 2 (acac) 2 ] in methanol yielded the mixed ligand complexes bearing a solvent molecule as co-ligand, [MoO 2 (L)MeOH)]. The ligands and complexes were characterized by using analytical and spectroscopic methods. As a representative sample, the dioxomolybdenum(VI) complex of the N4-butyl-substituted thiosemicarbazone was crystallographically examined to ensure the expected structures. X-ray data showed a distorted octahedral environment of molybdenum center. Tyrosinase, collagenase inhibition and antioxidant properties of the compounds were investigated using spectroscopic methods. The ligand and complexes exhibited the different inhibitory activities depending on N-alkyl substituents. Similarly, also the antioxidant capacity of the compounds changed in relation to the substituents. Image 1 • Four novel dioxomolybdenum complexes of thiosemicarbazone were prepared and characterized. • Antioxidant, antityrosinase, and anticollagenase activities of all compounds were firstly investigated depending on N-alkyl substituents. [ABSTRACT FROM AUTHOR]

Graphical abstract Abstract The extremely complex tumor microenvironment (TME) in humans is the major responsible for the therapeutic failure in cancer nanomedicine. A new concept of disease-driven nanomedicine, henceforth named "Theranomics", which attempts to target cancer cells and TME on the whole, represents an attractive alternative. Herein, a nanomedicine able to co-deliver doxorubicin and a tumor suppressive proteolytic protein such as collagenase-2 was developed. We successfully obtained superparamagnetic nanogels (SPIONs/Doco@Col) via the intermolecular azide-alkyne Huisgen cycloaddition. We demonstrated that a local ECM degradation and remodeling in solid tumors by means of collagenase-2 could enhance tumor penetration of nanomedicines and the in situ sustained release of the drug payload throughout 3-D tumor spheroids up to the core (parenchyma), thus enabling a synergistic and efficient anticancer effect toward highly invasive breast tumors. We illustrate that SPIONs/Doxo@Col is also capable of reducing the invasivity of cancer cells. [ABSTRACT FROM AUTHOR]

Highlights • Dental resin restoratives face biodegradative challenges from host- and bacterial- factors. • Degradation at the restoration-tooth margin contributes to secondary caries. • Degradation by-products increase degradative factors activity and bacterial virulence. • Reproducing biodegradative effects in vitro is vital to assessing new materials. • New materials should inhibit biodegradation as well as inhibit bacterial proliferation. Abstract Objectives Despite decades of development and their status as the restorative material of choice for dentists, resin composite restoratives and adhesives exhibit a number of shortcomings that limit their long-term survival in the oral cavity. Herein we review past and current work to understand these challenges and approaches to improve dental materials and extend restoration service life. Methods Peer-reviewed work from a number of researchers as well as our own are summarized and analyzed. We also include yet-unpublished work of our own. Challenges to dental materials, methods to assess new materials, and recent material improvements and research directions are presented. Results Mechanical stress, host- and bacterial-biodegradation, and secondary caries formation all contribute to restoration failure. In particular, several host- and bacterial-derived enzymes degrade the resin and collagen components of the hybrid layer, expanding the marginal gap and increasing access to bacteria and saliva. Furthermore, the virulence of cariogenic bacteria is up-regulated by resin biodegradation by-products, creating a positive feedback loop that increases biodegradation. These factors work synergistically to degrade the restoration margin, leading to secondary caries and restoration failure. Significant progress has been made to produce hydrolytically stable resins to resist biodegradation, as well as antimicrobial materials to reduce bacterial load around the restoration. Ideally, these two approaches should be combined in a holistic approach to restoration preservation. Significance The oral cavity is a complex environment that poses an array of challenges to long-term material success; materials testing conditions should be comprehensive and closely mimic pathogenic oral conditions. [ABSTRACT FROM AUTHOR]

Abstract Developing effective therapeutics for osteoarthritis (OA) necessitates that such molecules can reach and target chondrocytes within articular cartilage. However, predicting how well very large therapeutic molecules diffuse through cartilage is often difficult, and the relationship between local transport mechanics for these molecules and tissue heterogeneities in the tissue is still unclear. In this study, a 150 kDa antibody diffused through the articular surface of healthy and enzymatically degraded cartilage, which enabled the calculation of local diffusion mechanics in tissue with large compositional variations. Local cartilage composition and structure was quantified with Fourier transform infrared (FTIR) spectroscopy and second harmonic generation (SHG) imaging techniques. Overall, both local concentrations of aggrecan and collagen were correlated to local diffusivities for both healthy and surface-degraded samples (0.3 > R2 < 0.9). However, samples that underwent surface degradation by collagenase exhibited stronger correlations (R2 > 0.75) compared to healthy samples (R2 < 0.46), suggesting that the highly aligned collagen at the surface of cartilage acts as a barrier to macromolecular transport. [ABSTRACT FROM AUTHOR]

There appears to be controversy regarding differing patient and physician perceptions of adverse effects (AEs) in the treatment of Dupuytren disease with collagenase clostridium histolyticum (CCH). The aim of this study was to compare the number, type, and severity of AEs perceived and reported by patients and by their physician To assess AEs following CCH injection in a standardized way, patients were given a list of predefined complications and asked to rate their severity on a 4-point Likert scale ranging from 1 (serious) to 4 (insignificant). Eighty-five patients were included. Patients reported fewer AEs than their physician (mean, 1.48 vs 2.18). There was no agreement between physician- and patient-reported AEs except for skin lacerations, which showed fair agreement (κ = 0.257). Patients and physicians differ in their evaluation of AEs due to CCH treatment in Dupuytren disease. A fair level of agreement was observed for skin lacerations. Greater consensus is needed when defining AEs associated with CCH in the treatment of Dupuytren disease. [ABSTRACT FROM AUTHOR]

Matrix metalloproteinases (MMPs) are responsible for the turnover of intramuscular connective tissue in live animals. We hypothesize that MMPs may play a role in postmortem aging of beef muscles for the degradation of connective tissues. Four different experiments were performed to: 1) characterize MMP activity during postmortem aging of beef; 2) determine if the native beef MMP can contribute to connective tissue degradation in a simulated standard industry postmortem aging condition; 3) explore approaches to improve the native beef MMP activity and 4) characterize MMP activity in beef from cattle supplemented with supranutritional level of Zn. In experiment 1, MMP was active throughout the entire aging periods (3, 21, 42 and 63 d) for beef muscles Longissimus lumborum , Gluteus medius and Gastrocnemius , and the unknown MMP responsible for the collagen degradation was identified as MMP-9 by Western Blot. In experiment 2 and 3, MMP-9 activity was noticeable in the gels after 42 d of storage in the cooler. Moreover, the addition of ZnCl 2 in the model system significantly increased MMP-9 activity when compared to the control (P < 0.01). In experiment 4, Longissimus thoracis from animals supplemented with a supranutritional Zn level had increased Zn availability and MMP-9 activity than those from animals fed with a control diet (P < 0.05). Further research is needed better understand MMP-9 mechanism during postmortem aging of meat. With a better understanding of MMP-9 in the aging process, the beef industry can provide better connective tissue management strategies for lower-quality beef cuts. [ABSTRACT FROM AUTHOR]

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Collagen fibre remodelling is a strain dependent process which is stimulated by the degradation of existing collagen. To date, literature has focussed on strain dependent degradation of pure collagen or structurally simple collagenous tissues, often overlooking degradation within more complex, heterogenous soft tissues. The aim of this study is to identify, for the first time, the strain dependent degradation behaviour and mechanical factors influencing collagen degradation in arterial tissue using a combined experimental and numerical approach. To achieve this, structural analysis was carried out using small angle light scattering to determine the fibre level response due to strain induced degradation. Next, strain dependent degradation rates were determined from stress relaxation experiments in the presence of crude and purified collagenase to determine the tissue level degradation response. Finally, a 1D theoretical model was developed, incorporating matrix stiffness and a gradient of collagen fibre crimp to decouple the mechanism behind strain dependent arterial degradation. SALS structural analysis identified a strain mediated degradation response in arterial tissue at the fibre level not dissimilar to that found in literature for pure collagen. Interestingly, two distinctly different strain mediated degradation responses were identified experimentally at the tissue level, not seen in other collagenous tissues. Our model was able to accurately predict these experimental findings, but only once the load bearing matrix, its degradation response and the gradient of collagen fibre crimp across the arterial wall were incorporated. These findings highlight the critical role that the various tissue constituents play in the degradation response of arterial tissue. Statement of Significance Collagen fibre architecture is the dominant load bearing component of arterial tissue. Remodelling of this architecture is a strain dependent process stimulated by the degradation of existing collagen. Despite this, degradation of arterial tissue and in particular, arterial collagen, is not fully understood or studied. In the current study, we identified for the first time, the strain dependent degradation response of arterial tissue, which has not been observed in other collagenous tissues in literature. We hypothesised that this unique degradation response was due to the complex structure observed in arterial tissue. Based on this hypothesis, we developed a novel numerical model capable of explaining this unique degradation response which may provide critical insights into disease development and aid in the design of interventional medical devices. [ABSTRACT FROM AUTHOR]

Fibrosis is a common lesion in different pathologic diseases and defined by the excessive accumulation of collagen. Different approaches have been used to treat different conditions characterized by fibrosis. The FDA and EMA approved the use of collagenase to treat palmar fibromatosis (Dupuytren’s contracture). The EMA approved additionally its use in severe Peyronie’s disease, but it has been used off label in other conditions [1,2]. The approved treatment includes up to three (in palmar fibromatosis) or up to eight (in penile fibromatosis) injections followed by finger extension or penile modeling procedures, typically causing severe pain. Frequent single injections are adequate to treat palmar fibromatosis [3]. The need to repeatedly inject doses of this enzyme can be due to the labile nature of collagenase, which exhibits a complete activity loss after a short period of time. This study presents a novel strategy to manage this enzyme based on the synthesis of polymeric nanocapsules that contain collagenase encapsulated within their matrix. These nanocapsules have been engineered for achieving a gradual release of the encapsulated enzyme for a longer time, which can be up to ten days. The efficacy of these nanocapsules has been tested in a murine model of local dermal fibrosis, and the results demonstrate a reduction in fibrosis greater than that with the injection of free enzyme; this type of treatment showed a significant improvement compared to conventional therapy of free collagenase. Statement of Significance The use of proteins as therapeutic molecules has recently attracted great interest. Collagenase injection is the current treatment for fibrotic diseases. Unfortunately, proteins have a low stability and presume several repetition cycles to obtain an effective treatment. This article describes a novel treatment for these types of diseases using collagenase nanocapsules designed to exhibit a sustainable release of the encapsulated enzyme, which maintains the enzymatic activity for a long period of time. The therapeutic effect of nanocapsules was tested in a murine mouse model of local dermal fibrosis, and the results showed an important improved effect compared to the effect of the administration of free enzyme. These results indicate a high potential for this novel system to improve the current treatment for fibrotic diseases. [ABSTRACT FROM AUTHOR]

The primary pathogen of the Great Barrier Reef sponge Rhopaloeides odorabile , recently identified as a novel strain (NW4327) of Pseudoalteromonas agarivorans , produced collagenase which degraded R. odorabile skeletal fibers. We now report the collagenase of P. agarivorans as a metalloprotease which required Ca 2+ and Zn 2+ as cofactors. The collagenase was a TonB dependent receptor (TBDR) having a carboxypeptidase regulatory like domain (CRLD) in the N-terminal along with an outer membrane (OM) channel superfamily domain. The genes for TBDR sub-components and collagenase formed one unified entity in the genome of P. agarivorans NW4327. This association of a collagenase with a TBDR distinguished it from all known functional collagenases till date and for the first time, established the enzymatic capability of TBDRs. Predicted TBDR model demonstrated only 15% identity with ferripyoverdin receptor and the CRLD displayed merely 24% identity with carboxypeptidase catalytic chain. Presence of signal peptide, lack of transmembrane helices, absence of N-terminal in the cytoplasmic side, extracellular localization and recovery from the culture supernatant implicated that the TBDR was secreted. Stronger binding of the collagenase with marine sponge type IV collagen than type I collagen, revealed through molecular docking, indicated higher specificity of the enzyme towards type IV collagen. [ABSTRACT FROM AUTHOR]

Collagenase is specific protease which capable to hydrolyze the native triple helical or denatured collagen. It is a promising enzyme in view of their potential industrial and biomedical applications. Bacterial collagenases may differ significantly owing to their structure and functional characteristics. The process of collagen hydrolysis by collagenase is still unresolved due to imprecise collagenase structural characteristics. Therefore, the present study aimed to understand the molecular, structural, and functional characteristics of collagenase from Pseudomonas aeruginosa using various computational approaches. The physico-chemical features and secondary structure of the collagenase have been explored. The three-dimensional (3D) model of collagenase has been constructed and verified by various tools such as RAMPAGE, verify 3D, and ERRAT. The presence of more than 90% residues in favored region of Ramachandran plot revealed the good quality of protein model. The result of verify3D tool also indicates that 3D structural modeling of the selected collagenase protein is acceptable. The present study provides a detailed overview of the molecular, functional, and structural characteristics of collagenase from P. aeruginosa which might also be helpful to understand the possible structure and functions of unknown proteins. [ABSTRACT FROM AUTHOR]

Objectives To evaluate the effect of mussel adhesive protein (MAP) on collagenase activity, dentin collagen degradation and microtensile dentin bond strength (μTBS). Methods Three groups were designed: 1. experimental group: treated with MAP; 2. positive control: treated with GM6001 (collagenase-inhibitor); 3. negative control: treated with distilled water (DW). For collagenase activity, Clostridiopeptidase-A was added to each group (n = 5), and collagenase activity was assessed by colorimetric assay. For dentin collagen degradation, thirty dentin slabs were allocated to the three above groups (n = 10). Dentin collagen degradation was evaluated by measuring released hydroxyproline by colorimetric assay after being incubated in Clostridiopeptidase-A for 7 days. For microtensile bond strength, sixty human third molars with flat dentin surfaces were etched by phosphoric acid and then assigned to the three above groups (n = 20). An etch-and-rinse adhesive system was applied to all three groups as stated in standard clinic protocol. The test of μTBS was performed before and after thermocycling and collagenase challenge. Results The collagenase activities (nmol/min/mg) in the group of MAP was significantly less inactive compared to the group of GM6001 and DW (MAP < GM6000 < DW, p < 0.01). The hydroxyproline concentrations (μg/mL) was significantly less in the group of MAP compared to the group of GM6001 and DW (MAP < GM6000 < DW, p < 0.01). While there was no significant difference in the immediate μTBS (MPa) among three groups (p > 0.06), the value of μTBSs after thermocycling and collagenase challenge was significantly greater in the group of MAP and GM6001 compared to the group of DW (MAP, GM6000 > DW, p < 0.001). Significance MAP inhibits collagenase activity, prevents dentin collagen degradation, and delays the deterioration of the dentin bonding of composite restoration over time. [ABSTRACT FROM AUTHOR]

The search for natural principles is attracting much interest in the field of skin care. Fabaceae are major agricultural crops and potential sources of bioactive compounds with possible applications in human health and skin care. This study concerns the biological activities of the legume crops Melilotus officinalis (L.) Pall. and Lespedeza capitata Michx. for their potential use in skin care applications. The effects of plant ethanolic extracts at doses ranging from 0.25 to 50 μg/mL (from 1 to 5000 μg/mL in cell viability assays) were evaluated using in vitro tests on HaCaT human keratinocytes, 46BR.1N fibroblasts, and adipocyte cell cultures, and on matrix-degrading enzymes. MTT assay revealed weak effects on cell viability (IC 50 > 1000 μg/mL) and significant increase of fibroblast growth rate with both extracts. Similar induction of cell motility by the two extracts was observed on keratinocytes, while on fibroblasts M. officinalis induced a stronger effect with respect to L. capitata . Cell-free enzymatic assays showed stronger collagenase inhibition by L. capitata , while an ELISA assay revealed more efficient stimulation of fibroblast collagen production by M. officinalis . Oil-Red-O adipocyte staining showed more pronounced lipolytic effect of M. officinalis with respect to L. capitata . Both extracts showed the ability of stimulating skin cells in order to promote tissue regeneration, prevent skin aging, and reduce fat deposition. In most cases, different patterns of activation/inhibition were observed. Data indicate that these legume crops could be profitably exploited in skin care applications, possibly in combined formulations for the development of antiaging and anticellulite products. [ABSTRACT FROM AUTHOR]

Biomimetic mineralization emphasizes reversing the process of dental caries through bio-inspired strategies, in which mineralization promotion and collagen protection are equally important. In this study, carboxymethyl chitosan (CMC) was deemed as an analog of glycosaminoglycan for biomimetic modification of collagen, both of the mineralization facilitation and collagen protection effect were evaluated. Experiments were carried out simultaneously on two-dimensional monolayer reconstituted collagen model, three-dimensional reconstituted collagen model and demineralized dentin model. In three models, CMC was successfully cross-linked onto collagen utilizing biocompatible 1-Ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy sulfosuccinimide sodium salt to achieve biomodification. Results showed that CMC biomodification increased collagen's hydrophilicity, calcium absorption capacity and thermal degradation resistance. In demineralized dentin model, the activity of endogenous matrix metalloproteinases was significantly inhibited by CMC biomodification. Furthermore, CMC biomodification significantly improved cross-linking and intrafibrillar mineralization of collagen, especially in the two-dimensional monolayer reconstituted collagen model. This study provided a biomimetic mineralization strategy with comprehensive consideration of collagen protection, and enriched the application of chitosan-based materials in dentistry. • Carboxymethyl chitosan was deemed as an analog of glycosaminoglycan for chemical biomodification of collagen models. • Three kinds of collagen models with gradient complexity were explored simultaneously. • Both of the mineralization facilitation and collagen protection effect were evaluated. [ABSTRACT FROM AUTHOR]

Sonodynamic therapy (SDT), a combination of low-intensity ultrasound with a sonosensitizer, has been explored as a promising alternative for cancer therapy. However, condensed extracellular matrix (ECM) resulting in poor perfusion and extreme hypoxia in solid tumor potentially compromises effective SDT. Herein, we develop a novel cleavable collagenase-assistant and O 2 -supplied nanosonosensitizer (FePO 2 @HC), which is embedded through fusing collagenase (CLG) and human serum albumin (HSA), followed by encapsulating Ferric protoporphyrin (FeP) and dioxygen. As a smart carrier, HSA is stimuli-responsive and collapsed by reduced glutathione (GSH) overexpressed in tumor, resulting to the release of the components in FePO 2 @HC. The released CLG acting as an artificial scissor, degrades the collagen fibers in tumor, thus, breaking tumor tissue and enhancing FePO 2 accumulation in tumor inner with higher than that without CLG. Simultaneously, oxygen molecules are released from FePO 2 in hypoxic environment and alleviate the tumor hypoxia. As a sonosensitizer, FeP is subsequently irradiated by ultrosound wave (US) and activates surrounding dioxygen to generate amount of singlet oxygen (1O 2). Contributed from the ECM-degradation, such SDT-based nanosystem with increased sonosensitizer permeability and oxygen content highly improved the tumor inhibition efficacy without toxic effects. This study presents a new paradigm for ECM depletion-based strategy of deep-seated penetration, and will expand the nanomedicine application of metalloporphyrin sonosensitizers in SDT. [ABSTRACT FROM AUTHOR]

Saffron, Crocus sativus L, is a perennial spice herb. It has been extensively studied for its antioxidant, antidepressant, and anti-inflammatory properties, and has recently gained new interest for use in high-end cosmetics. The present work aims to elucidate the skin-protective properties of saffron in human dermal fibroblasts. The skin-protective properties of saffron extract were evaluated in terms of tyrosinase and collagenase inhibition activities, antioxidant activity in mouse macrophage cells, collagen synthesis and hyaluronic acid synthesis and cell migration activity in primary dermal fibroblast normal human neonatal cells (HDFn). Saffron's main phytoactive constituents - crocins, picrocrocin, safranal, and crocetin - were quantified by LC-MS at 91.0, 61.5, 3.6, and 1.9 mg/g, respectively. Saffron extract inhibited tyrosinase and collagenase with IC 50 0.78 mg/mL and 0.1 mg/mL, respectively. Saffron extracts (100–200 μg/mg) suppressed reactive oxygen species and nitric oxide generation in lipopolysaccharide-stimulated RAW 264.7 murine macrophage cells and promoted collagen and hyaluronic acid synthesis in HDFn. Saffron extract at 25 μg/mg significantly promoted migration of HDFn cells (wound healing capacity) compared to control (no treatment). The findings highlighted the potential benefits of saffron extract relevant to skin resilience. [Display omitted] • Skin-protective properties of saffron were elucidated in human dermal fibroblasts. • Saffron extract exhibited tyrosinase and collagenase inhibition activities. • Saffron extract promoted collagen and hyaluronic acid synthesis in HDFn. • Picrocrocin and crocetin stimulated cell migration to a greater degree than safranal in HDFn. [ABSTRACT FROM AUTHOR]

• Specific nontoxic fractions from Cosmos caudatus (CC) exerts antioxidant properties. • These fractions inhibit collagenase, MMP-1 & MMP-3 expressions in CCD-966SK cells. • 14 compounds are identified from CC fractions with skin anti-aging properties. • CC chemical fingerprint is established by quantifying 8 major & bioactive markers. • Flavonoids comprised CC fractions synergistically suppressed MMP-3 expression. Skin aging is associated with degradation of collagen by matrix metalloproteinases (MMPs), which leads to loss of skin elasticity and formation of wrinkles. Cosmos caudatus Kunth (CC) has been traditionally claimed as an anti-aging agent in Malaysia. Despite its well-known antioxidant activity, the anti-aging properties of CC was not validated. This study aimed to investigate the anti-aging potential of CC extracts and fractions, particularly their inhibition of collagenase, MMP-1 and MMP-3 activities in human dermal fibroblasts CCD-966SK, followed by isolation, identification and analysis of their bioactive constituents. DPPH assay was firstly used to evaluate the antioxidant activity throughout the bioactivity-guided fractionation. Cell viability was determined using MTS assay. Collagenase activity was examined, while MMP-1 and MMP-3 expression were measured using qRT-PCR and western blotting. Then, chemical identification of pure compounds isolated from CC fractions was done by using ESIMS, 1H and 13C NMR spectroscopies. HPLC analyses were carried out for bioactive fractions to quantify the major components. Throughout the antioxidant activity-guided fractionation, fractions CC-E2 and CC-E3 with antioxidant activity and no toxicity towards CCD-966SK cells were obtained from CC 75% ethanol partitioned layer (CC-E). Both fractions inhibited collagenase activity, MMP-1 and MMP-3 mRNA and protein expression, as well as NF-κB activation induced by TNF-α in CCD-966SK cells. 14 compounds, which mainly consists of flavonoids and their glycosides, were isolated. Quercitrin (14.79% w/w) and quercetin (11.20% w/w) were major compounds in CC-E2 and CC-E3, respectively, as quantified by HPLC. Interestingly, both fractions also inhibited the MMP-3 protein expression synergistically, compared with treatment alone. The quantified CC fractions rich in flavonoid glycosides exhibited skin anti-aging effects via the inhibition of collagenase, MMP-1 and MMP-3 activities, probably through NF-κB pathway. This is the first study reported on MMP-1 and MMP-3 inhibitory activity of CC with its chemical profile, which revealed its potential to be developed as anti-aging products in the future. [Display omitted] [ABSTRACT FROM AUTHOR]