Porcine Skin Collagen Research Articles

Recognition of MCF-7 breast cancer cells using native collagen probes: Collagen source effect

Developing superior cancer cell recognition probes is crucial for the development of tumor therapy and cancer early screening materials. In this study, we first achieved effective recognition of MCF-7 breast cancer cells using natural collagen probes. Through cell adhesion, cancer cell selective capture, and flow cytometry techniques, the binding efficiency of mammalian-derived collagens (bovine Achilles tendon collagen, porcine skin collagen) and fish-derived collagens (turbot skin collagen, grass carp skin collagen, mandarin fish skin collagen) to cancer cells (MCF-7 breast cancer cells) and normal cells (human umbilical vein endothelial cells, HUVECs) was analyzed and compared. The feasibility of different source collagens as probes for recognition of MCF-7 cells was explored in vitro. The results indicated that mammalian-derived collagens had a superior advantage over fish-derived collagens in recognizing MCF-7 cells, with bovine Achilles tendon collagen achieving a capture rate of up to 64.7 % in a multicellular co-culture system. Furthermore, in vivo imaging of BALB/c tumor-bearing mice confirmed the high-efficiency targeted recognition performance of the bovine Achilles tendon collagen probe for MCF-7 cells.

Assessment of Tilapia Skin Collagen for Biomedical Research Applications in Comparison with Mammalian Collagen.

Collagen is an important material for biomedical research, but using mammalian tissue-derived collagen carries the risk of zoonotic disease transmission. Marine organisms, such as farmed tilapia, have emerged as a safe alternative source of collagen for biomedical research. However, the tilapia collagen products for biomedical research are rare, and their biological functions remain largely unexamined. In this study, we characterized a commercial tilapia skin collagen using SDS-PAGE and fibril formation assays and evaluated its effects on skin fibroblast adhesion, proliferation, and migration, comparing it with commercial collagen from rat tails, porcine skin, and bovine skin. The results showed that tilapia skin collagen is a type I collagen, similar to rat tail collagen, and has a faster fibril formation rate and better-promoting effects on cell migration than porcine and bovine skin collagen. We also confirmed its application in a 3D culture for kidney cells' spherical cyst formation, fibroblast-induced gel contraction, and tumor spheroid interfacial invasion. Furthermore, we demonstrated that the freeze-dried tilapia skin collagen scaffold improved wound closure in a mouse excisional wound model, similar to commercial porcine or bovine collagen wound dressings. In conclusion, tilapia skin collagen is an ideal biomaterial for biomedical research.

A double-crosslinked nanocellulose-reinforced dexamethasone-loaded collagen hydrogel for corneal application and sustained anti-inflammatory activity

In cases of blinding disease or trauma, hydrogels have been proposed as scaffolds for corneal regeneration and vehicles for ocular drug delivery. Restoration of corneal transparency, augmenting a thin cornea and postoperative drug delivery are particularly challenging in resource-limited regions where drug availability and patient compliance may be suboptimal. Here, we report a bioengineered hydrogel based on porcine skin collagen as an alternative to human donor corneal tissue for applications where long-term stability of the hydrogel is required. The hydrogel is reinforced with cellulose nanofibers extracted from the Ciona intestinalis sea invertebrate followed by double chemical and photochemical crosslinking. The hydrogel is additionally loaded with dexamethasone to provide sustained anti-inflammatory activity. The reinforced double-crosslinked hydrogel after drug loading maintained high optical transparency with significantly improved mechanical characteristics compared to non-reinforced hydrogels, while supporting a gradual sustained drug release for 60 days in vitro. Dexamethasone, after exposure to crosslinking and sterilization procedures used in hydrogel production, inhibited tube formation and cell migration of TNFα-stimulated vascular endothelial cells. The drug-loaded hydrogels suppressed key pro-inflammatory cytokines CCL2 and CXCL5 in TNFα-stimulated human corneal epithelial cells. Eight weeks after intra-stromal implantation in the cornea of 12 New-Zealand white rabbits subjected to an inflammatory suture stimulus, the dexamethasone-releasing hydrogels suppressed TNFα, MMP-9, and leukocyte and fibroblast cell invasion, resulting in reduced corneal haze, sustained corneal thickness and stromal morphology, and reduced overall vessel invasion. This collagen-nanocellulose double-crosslinked hydrogel can be implanted to treat corneal stromal disease while suppressing inflammation and maintaining transparency after corneal transplantation. Statement of significanceTo treat blinding diseases, hydrogel scaffolds have been proposed to facilitate corneal restoration and ocular drug delivery. Here, we improve on a clinically tested collagen-based scaffold to improve mechanical robustness and enzymatic resistance by incorporating sustainably sourced nanocellulose and dual chemical-photochemical crosslinking to reinforce the scaffold, while simultaneously achieving sustained release of an incorporated anti-inflammatory drug, dexamethasone. Evaluated in the context of a corneal disease model with inflammation, the drug-releasing nanocellulose-reinforced collagen scaffold maintained the cornea's transparency and resisted degradation while suppressing inflammation postoperatively. This biomaterial could therefore potentially be applied in a wider range of sight-threatening diseases, overcoming suboptimal administration of postoperative medications to maintain hydrogel integrity and good vision.

Preparation of injectable porcine skin-derived collagen and its application in delaying skin aging by promoting the adhesion and chemotaxis of skin fibroblasts

Collagen, as the main component of human skin, plays a vital role in maintaining dermal integrity. Its loss will lead to dermis destruction and collapse, resulting in skin aging. At present, injection of exogenous collagen is an important means to delay skin aging. In this study, high-purity collagen was extracted from porcine skin. Our research revealed that it can effectively promote the adhesion and chemotaxis of HSF cells. It can also reduce the expression of β-galactosidase, decrease ROS levels, and increase the expression of the collagen precursors, p53 and p16 in HSF cells during senescence. After local injection into the aging skin of rats, it was found that the number of cells and type I collagen fibers in the dermis increased significantly, and the arrangement of these fibers became more uniform and orderly. Moreover, the important thing is that it is biocompatible. To sum up, the porcine skin collagen we extracted is an anti-aging biomaterial with application potential.

Identification of a highly stable bioactive 3-hydroxyproline-containing tripeptide in human blood after collagen hydrolysate ingestion

There are increasing reports demonstrating high bioavailability of 4-hydroxyproline (4Hyp)-containing oligopeptides after oral ingestion of collagen hydrolysate and their bioactivity. In contrast, no study investigates the fate of another collagen-specific but minor amino acid, 3Hyp. Here, we identified Gly-3Hyp-4Hyp tripeptide in human blood at high concentrations, comparable to other 4Hyp-containing oligopeptides, after ingesting porcine skin collagen hydrolysate. Additionally, Gly-3Hyp-4Hyp uniquely maintained the maximum concentration until 4 h after the ingestion due to its exceptionally high resistance to peptidase/protease demonstrated by incubation with mouse plasma. In mice, oral administration of collagen hydrolysate prepared from bovine tendon, which contains a higher amount of 3Hyp, further increased blood Gly-3Hyp-4Hyp levels compared to that from bovine skin. Furthermore, Gly-3Hyp-4Hyp showed chemotactic activity on skin fibroblasts and promoted osteoblast differentiation. These results highlight the specific nature of the Gly-3Hyp-4Hyp tripeptide and its potential for health promotion and disease treatment.

Cryogenically structured gelatin-based hydrogel as a resorbable macroporous matrix for biomedical technologies

Objective: to investigate the biological properties of a matrix made of cryogenically structured hydrogel in the form of a macroporous gelatin sponge, as well as the possibility of creating cell-engineered constructs (CECs) on its basis. Materials and methods. The main components of the cryogenically structured hydrogel were gelatin (type A) obtained from porcine skin collagen, N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide, (EDC) and urea (all from Sigma-Aldrich, USA). Surface morphology was examined using scanning electron microscopy (SEM). The degree of swelling in water of the samples was determined by gravimetric method. Cytotoxicity was studied on NIH3T3, a fibroblast cell line isolated from a mouse, and on human adipose-derived mesenchymal stem/stromal cells (hAMSCs) using IncuCyte ZOOM (EssenBioscience, USA). The metabolic activity of hAMSCs was assessed using PrestoBlue™ reagents (Invitrogen™, USA). To create CECs, we used hAMSCs, human hepatocellular carcinoma cell line HepG2 or human umbilical vein endothelial cell lines EA.hy926. Albumin content in the culture medium was determined by enzyme immunoassay. Ammonia metabolism rate was assessed after 90 minutes of incubation with 1 mM ammonium chloride (Sigma-Aldrich, USA) diluted in a culture medium on day 15 of the experiment. Results. Obtaining a cryogenically structured hydrogel scaffold in the form of macroporous gelatin sponge included freezing an aqueous solution of a gelatin+urea mixture, removal of polycrystals of frozen solvent by lyophilization, extraction of urea with ethanol and treatment of the cryostructurate with an ethanol solution of EDC. Scanning electron microscopy identified three types of pores on the carrier surface: large (109 ± 17 μm), medium (39 ± 10 μm), and small (16 ± 6 μm). The degree of swelling in water of the matrix samples was 3.8 ± 0.2 g H2O per 1 g of dry polymer. The macroporous gelatin sponge as a part of CEC was found to have the ability to support adhesion and proliferation of hAMSCs, EA.hy926 and HepG2 for 28, 15 and 9 days, respectively. Albumin secretion and ammonia metabolism when HepG2 cells were cultured on the gelatin sponge were detected. Conclusion. The use of a matrix made from macroporous cryogenically structured gelatin-based hydrogel for tissue engineering products is shown to be promising using a cell-engineered liver construct as a case.

Structural, functional, rheological, and biological properties of the swim bladder collagen extracted from grass carp (Ctenopharyngodon idella)

The objective of this study was to develop an aquatic collagen with high thermal stability as a possible alternative to terrestrial sources. Swim bladder collagen (SBC) was extracted from grass carp with high yield of 39.2% (db), and characterized as type I collagen. X-ray diffraction indicated that SBC maintains a native triple helical collagen structure. Secondary structure analysis revealed that SBC consists of 41.93% α-helices, 45.31% β-sheets, 12.76% β-turns, and 0% random coils. Proteins from SBC exhibited foaming and emulsifying superiority to proteins from terrestrial sources, such as chicken feet collagen. The thermal denaturation temperature of SBC (34.3 °C) is similar to porcine skin collagen. Dynamic frequency sweep tests showed that elasticity plays a dominant role in the SBC solution system as the concentration increases. The SBC solution system shows viscous behavior when the temperature is below Td. In addition, the steady shear tests showed that all of the SBC solutions exhibited pseudoplasticity with shear-thinning behavior. SBC could provide a suitable environment for MC3T3-E1 cell growth and maintain normal cellular morphology. Overall, the results indicated that SBC might has potential for further applications in food, cosmetics and biomedical fields.

Effect of cooking temperature on texture and flavour binding of braised sauce porcine skin

SummaryTo investigate the effect of cooking temperature (55, 65, 75, 85 and 95 °C) on texture and flavour binding of braised sauce porcine skin (BSPS), sensory acceptance, microstructure and flavour‐binding capacity were investigated during the processing of BSPS. Samples cooked at 85 and 95 °C showed better texture and aroma scores. Hardness and chewiness of BSPS were obviously improved at 85 and 95 °C than control group. Collagen structure was significantly destroyed over 85 °C. The porcine skin collagen heated at 85 and 95 °C showed relatively higher flavour‐binding capacity than other samples. The improvement of texture of BSPS was mainly attributed to the degradation of collagen. Higher aroma scores of BSPS were related to intense binding abilities with aroma compounds at 85 and 95 °C. Cooking at 85 or 95 °C could be an optimal cooking temperature for BSPS.

Purification, Characterization, and Application for Preparation of Antioxidant Peptides of Extracellular Protease from Pseudoalteromonas sp. H2.

The study reported on the isolation of a metalloprotease named EH2 from Pseudoalteromonas sp. H2. EH2 maintained more than 80% activity over a wide pH range of 5–10, and the stability was also nearly independent of pH. Over 65% activity was detected at a wide temperature range of 20–70 °C. The high stability of the protease in the presence of different surfactants and oxidizing agents was also observed. Moreover, we also investigated the antioxidant activities of the hydrolysates generated from porcine and salmon skin collagen by EH2. The results showed that salmon skin collagen hydrolysates demonstrated higher DPPH (1,1-diphenyl-2-picrylhydrazyl) (42.88% ± 1.85) and hydroxyl radical (61.83% ± 3.05) scavenging activity than porcine skin collagen. For oxygen radical absorbance capacity, the hydrolysates from porcine skin collagen had higher efficiency (7.72 ± 0.13 μmol·TE/μmol). Even 1 nM mixed peptides could effectively reduce the levels of intracellular reactive oxygen species. The two types of substrates exerted the best antioxidant activity when hydrolyzed for 3 h. The hydrolysis time and type of substrate exerted important effects on the antioxidant properties of hydrolysates. The hydrolyzed peptides from meat collagens by proteases have good antioxidant activity, which may have implications for the potential application of marine proteases in the biocatalysis industry.

Isolation of ice structuring collagen peptide by ice affinity adsorption, its ice-binding mechanism and breadmaking performance in frozen dough

Ice structuring collagen peptides (ISCPs) from porcine skin collagen hydrolysates were isolated by an affinity adsorption system, and then its antifreeze and cryoprotection activity, ice-binding mechanism and making performance of steamed bread were investigated. The results showed ISCPs with maximal thermal hysteresis activity (2.76°C) were obtained with a collagen hydrolysate concentration of 1 mg/mL, adsorption time of 10 hr and adsorption temperature of −5°C during the second round of adsorption. After purification, MS spectrum analyzed one of the ISCPs and inferred to be GLLGPLGPRGLL with a molecular weight of 1163.8 Da. The binding of ISCP to ice could be attributed to the structural match and hydrogen bond formation between ice surface and the ISCP. Compared with the control group, the baking and textural characteristics of frozen dough were significantly improved by ISCP at an addition concentration of 0.3% (wt/vol) after 4 weeks frozen storage. Practical applications Porcine skin represents a problematic by-product in meat manufacturing industry due to its high pollutant load, and it is usually discarded as waste or used for low value purposes. In order to find potential applications for this by-product, ISCPs from porcine skin hydrolysates were isolated by an affinity adsorption system. The result demonstrated that ISCPs exhibited a strong antifreeze and cryoprotection activity, and had a potential application in food preservation and cryo-protection of cells and tissues.

In Vitro Fabrication and Physicochemical Properties of a Hybrid Fibril from Xenogeneic Collagens

A Hybrid collagen fibril (HCF) assembled from xenogeneic collagens is a special kind of collagen fibrils in vivo and plays an important role in living systems. Inspired by nature, can a HCF form in vitro? Herein, we fabricated a new HCF by neutralizing a mixture of type I bullfrog (Rana catesbeiana Shaw) skin collagen and porcine (Sus scrofa domesticus) skin collagen with a phosphate buffer, and investigated its physicochemical properties. Self-assembly kinetics and fluorescence-quenching experiments showed that a significant intermolecular interaction and co-assembly behavior occurred between bullfrog skin collagen and porcine skin collagen, thus confirming that xenogeneic collagens can self-assemble to form HCF. Differential scanning calorimetry revealed that the thermal stability of HCF was completely different from that of the syngeneic bullfrog skin and porcine skin collagen fibrils. This finding indicated that a new kind of collagen fibril was fabricated successfully. Scanning electron microscopy and transmission electron microscopy tests showed that the diameters and D-periodicity lengths of HCF were smaller than those of the syngeneic collagen fibrils, suggesting that the morphological features of HCF were distinguished from those of the syngeneic fibril samples. Moreover, viscoelasticity of a collagen gel also changed after the self-assembly of xenogeneic collagens. Meanwhile, the obtained hybrid gel still exhibited good biocompatibility and cell proliferation properties. Finding from this work provides a new idea for the improvement or regulation of collagen-based products performance.

Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin.

Super- and sub-critical water treatments have been of interest as novel methods for protein hydrolysis. In the present study, we studied the effect of sub-critical water (Sub-H2O, 300℃, 80 bar) treatment as well as super-critical water (Super-H2O, 400℃, 280 bar) treatment on the physicochemical properties of porcine skin (PS), which has abundant collagen. Porcine skin was subjected to pre-thermal treatment by immersion in water at 70℃, and then treated with sub- or super-critical water. Physicochemical properties of the hydrolysates, such as molecular weight distribution, free amino acid content, amino acid profile, pH, color, and water content were determined. For the molecular weight distribution analysis, 1 kDa hydrolyzed porcine skin (H-PS) was produced by Super-H2O or Sub-H2O treatment. The free amino acid content was 57.18 mM and 30.13 mM after Sub-H2O and Super-H2O treatment, respectively. Determination of amino acid profile revealed that the content of Glu (22.5%) and Pro (30%) was higher after Super-H2O treatment than after Sub-H2O treatment, whereas the content of Gly (28%) and Ala (13.1%) was higher after Sub-H2O treatment. Super-H2O or Sub-H2O treatment affected the pH of PS, which changed from 7.29 (Raw) to 9.22 (after Sub-H2O treatment) and 9.49 (after Super-H2O treatment). Taken together, these results showed that Sub-H2O treatment was slightly more effective for hydrolysis than Super-H2O was. However, both Sub-H2O and Super-H2O treatments were effective processing methods for hydrolysis of PS collagen in a short time and can be regarded as a green chemistry technology.

Biocompatibility of Novel Type I Collagen Purified from Tilapia Fish Scale: An In Vitro Comparative Study.

Type I collagen (COL-1) is the prevailing component of the extracellular matrix in a number of tissues including skin, ligament, cartilage, bone, and dentin. It is the most widely used tissue-derived natural polymer. Currently, mammalian animals, including pig, cow, and rat, are the three major sources for purification of COL-1. To reduce the risk of zoonotic infectious diseases transmission, minimize the possibility of immunogenic reaction, and avoid problems related to religious issues, exploration of new sources (other than mammalian animals) for the purification of type I collagen is highly desirable. Hence, the purpose of the current study was to investigate the in vitro responses of MDPC-23 to type I collagen isolated from tilapia scale in terms of cellular proliferation, differentiation, and mineralization. The results suggested that tilapia scale collagen exhibited comparable biocompatibility to porcine skin collagen, indicating it might be a potential alternative to type I collagen from mammals in the application for tissue regeneration in oral-maxillofacial area.

Amino acid composition and antioxidant activities of hydrolysates and peptide fractions from porcine collagen

The amino acid composition and antioxidant activities of different hydrolysates from porcine collagen were analyzed. The gelatin was hydrolyzed for antioxidative peptides with various proteases, namely papain, protease from bovine pancreas, protease from Streptomyces, and cocktail mixture of protease from bovine pancreas and protease from Streptomyces. The hydrolysates were assessed using methods of DPPH radical-scavenging ability, metal-chelating ability and lipid peroxidation inhibition activity. It was found that the collagen hydrolysates by different protease treatments had different amino acid compositions and antioxidant properties. However, the contents of Hyp and Pro were improved and the content of Gly was decreased in each collagen hydrolysate compared with collagen. The hydrolysate prepared with the cocktail mixture of proteases, which exhibited the highest antioxidant activity, was separated into 6 fractions by gel filtration chromatography. Fraction 2 was further separated by ion exchange chromatography. Fraction 2b with abundant basic amino acids and Fraction 2d which was slightly acidic fractions had higher radical-scavenging and metal-chelating activities, and both Fraction 2b and 2d contained more hydrophobic amino acids. The results confirmed that the antioxidative peptides were rich in Hyp, Pro and Gly, which accounted for half of amino acid composition. This article added further support to the preparation of natural antioxidative peptides from porcine skin collagen.

Polymerization and matrix physical properties as important design considerations for soluble collagen formulations

Despite extensive use of type I collagen for research and medical applications, its fibril-forming or polymerization potential has yet to be fully defined and exploited. Here, we describe a type I collagen formulation that is acid solubilized from porcine skin collagen (PSC), quality controlled based upon polymerization potential, and well suited as a platform polymer for preparing three-dimensional (3D) culture systems and injectable/implantable in vivo cellular microenvironments in which both relevant biochemical and biophysical parameters can be precision-controlled. PSC is compared with three commercial collagens in terms of composition and purity as well as polymerization potential, which is described by kinetic parameters and fibril microstructure and mechanical properties of formed matrices. When subjected to identical polymerization conditions, PSC showed significantly decreased polymerization times compared to the other collagens and yielded matrices with the greatest mechanical integrity and broadest range of mechanical properties as characterized in oscillatory shear, uniaxial extension, and unconfined compression. Compositional and intrinsic viscosity analyses suggest that the enhanced polymerization potential of PSC may be attributed to its unique oligomer composition. Collectively, this work demonstrates the importance of standardizing next generation collagen formulations based upon polymerization potential and provides preliminary insight into the contribution of oligomers to collagen polymerization properties.

Antihypertensive Effect of Enzymatic Hydrolysate of Collagen and Gly-Pro in Spontaneously Hypertensive Rats

Continuous oral feeding of enzymatic hydrolysate of porcine skin collagen showed an antihypertensive effect in spontaneously hypertensive rats (SHRs). We isolated an angiotensin I-converting enzyme (ACE) inhibitory peptide, Gly-Phe-Hyp-Gly-Pro (IC(50)=91 microM), from the hydrolysate, but the ACE inhibitory activities of the other peptides isolated were weak. Although the ACE inhibitory activity of Gly-Pro (IC(50)=360 microM) was not potent, Gly-Pro exists in collagen as a large number of repeated sequences. We then examined the antihypertensive effect of Gly-Pro. Orally administered Gly-Pro at 500 mg/kg significantly decreased the blood pressure of SHRs, and at 50 mg/kg it also showed a tendency to lower the blood pressure. Oral administration of Gly-Phe-Hyp-Gly-Pro (10 or 30 mg/kg) also decreased the blood pressure of SHRs.

Genital Prolapse Repair Using Porcine Skin Implant and Bilateral Sacrospinous Fixation: Midterm Functional Outcome and Quality-of-Life Assessment

To evaluate the midterm anatomic and functional outcome of genital prolapse repair by the vaginal route using a porcine skin implant (Pelvicol) and bilateral sacrospinous fixation. From May 2001 to June 2006, 101 patients with Stage III-IV genital prolapse were treated using a porcine skin collagen implant and bilateral sacrospinous fixation. The functional results were evaluated using the Pelvic Floor Distress Inventory short form, Pelvic Organ Prolapse Distress Inventory-6, Colorectal Anal Distress Inventory-8, Urogenital Distress Inventory-6, Pelvic Floor Impact Questionnaire-7 (including Urinary Impact Questionnaire-7, Pelvic Organ Prolapse Impact Questionnaire-7, and Colo-Rectal-Anal Impact Questionnaire-7), and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire. Of the 101 patients, 89 (88%) completed all the questionnaires. The mean follow-up was 38 +/- 18 months. An improvement was noted in the Pelvic Organ Prolapse Distress Inventory-6 (P < .0001), Urogenital Distress Inventory-6 (P = .001), and Pelvic Floor Distress Inventory scores (P < .0001) but not in the Colorectal Anal Distress Inventory-8 scores. An improvement was noted in the Urinary Impact Questionnaire-7 (P < .0001), Pelvic Organ Prolapse Impact Questionnaire-7 (P < .0001), and Pelvic Floor Impact Questionnaire Short Form 7 (p < 0.0001) scores but not in the Colo-Rectal-Anal Impact Questionnaire-7 scores. The quartile distribution showed that women with a preoperative Pelvic Floor Distress Inventory-short form score >133 had a 45% chance of postoperative improvement and those with a preoperative Pelvic Floor Impact Questionnaire Short Form 7 score >195 had an 81% chance of postoperative improvement. Using multivariate regression analysis, the preoperative Pelvic Floor Distress Inventory score was negatively predictive of satisfaction (t = -2.03, P = .05) and the preoperative Pelvic Floor Impact Questionnaire Short Form 7 score was positively predictive of satisfaction (t = 2.40, P = .02). No differences in the pre- and postoperative Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire scores were noted. The results of our study have shown that biologic implantation and bilateral sacrospinous fixation are effective in high-grade genital prolapse repair. Validated questionnaires are potentially useful tools to predict the postoperative outcome.

Chirurgie du prolapsus : cœlioscopie ou voie basse ?

Objective To compare the safety and the efficacy of the laparoscopic and vaginal technique for the surgical management of pelvic organ prolapse, with systematic support of the three compartments and prosthetic reinforcements. Patients and methods Retrospective study of 154 patients presenting a stage 3 or 4 prolapse on one of the three compartments. Laparoscopic procedures were performed with subtotal hysterectomy, double synthetic prosthesis attached to promontory, and douglassectomy. Vaginal procedures were performed with vaginal hysterectomy, anterior colporrhaphy with a hammock using porcine skin collagen implant fixed by transobturator passages, unilateral sacral colpopexy and posterior colporrhaphy. Monitoring was performed at six months and then annually. Results The laparoscopic technique requires a more important operating time, but a shorter hospitalization. The discovery of three carcinoma reinforces the idea of the interest of uterine radical surgery in these patients. The tolerance of prostheses by laparoscopy is safe. The biological prostheses, introduced vaginally, offer the same advantages. The anatomical results in the medium term (30 months) seem more favorable to laparoscopy than transvaginal approach, as well as functional results but they still need to better evaluated. Discussion and conclusion The two techniques must coexist, ideally without competing with each other but rather complementarily, as the overall rate of recurrence, requiring additional procedure does not exceed 2%. It is therefore important that surgeons, who support prolapse, have a good comprehensive training of the laparoscopic and vaginal techniques.

Collagen from common minke whale (Balaenoptera acutorostrata) unesu

Collagen was prepared from common minke whale unesu and characterised. The yield of collagen was high, about 28.4% on a wet weight basis. By SDS–PAGE and CM-Toyopearl 650M column chromatography, the collagen was classified as type I collagen. The denaturation temperature of the collagen was 31.5°C, about 6–7°C lower than that of porcine collagen. Attenuated total reflectance-FTIR analysis indicated that acid-soluble collagen from common minke whale unesu held its triple helical structure well, but the structures of porcine skin collagen and pepsin-solubilized collagen from common minke whale unesu were changed slightly, due to the loss of N- and C-terminus domains.

Isolation and identification of antioxidative peptides from porcine collagen hydrolysate by consecutive chromatography and electrospray ionization–mass spectrometry

The porcine skin collagen was hydrolyzed by different protease treatments to obtain antioxidative peptides. The hydrolysate of collagen by cocktail mixture of protease bovine pancreas, protease Streptomyces and protease Bacillus spp. exhibited the highest antioxidant activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, metal chelating and in a linoleic acid peroxidation system induced by Fe 2+. And degree of hydrolysis highly affected the antioxidant properties of the hydrolysates. Four different peptides showing strong antioxidant activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration chromatography, ion-exchange chromatography and high-performance liquid chromatography. The molecular masses and amino acid sequences of the purified antioxidant peptides were determined using electrospray ionization (ESI) mass spectrometry. One of the antioxidative peptides, Gln-Gly-Ala-Arg, was then synthesized and the antioxidant activities measured using the aforementioned methods. The results confirmed the antioxidant activity of this peptide, and adds further support to its feasibility as a provider of natural antioxidants from porcine skin collagen protein.