Combining an HA + Cu (II) Site-Targeted Copper-Based Product with a Pruning Wound Protection Program to Prevent Infection with Lasiodiplodia spp. in Grapevine

The genus Lasiodiplodia has been reported from several grape growing regions and is considered as one of the fastest wood colonizers, causing Botryosphaeria dieback. The aim of this study was to (i) evaluate the efficacy of Esquive®, a biocontrol agent, on vineyard pruning wound protection, applied single or, in a combined protection strategy with a new site-targeted copper-based treatment (LC2017), and (ii) compare their efficacy with chemical protection provided by the commercially available product, Tessior®. For two seasons, protectants were applied onto pruning wounds, while LC2017 was applied throughout the season according to the manufacturer’s instructions. Pruning wounds of two different cultivars were inoculated with three isolates of Lasiodiplodia spp. Efficacy of the wound protectants, varied between both years of the assay and according to the cultivar studied but were able to control the pathogen to some extent. The application of LC2017 did not show clear evidence of improving the control obtained by the sole application of the other products tested. Nevertheless, LC2017 showed a fungistatic effect against Lasiodiplodia spp., in vitro, and has previously shown an elicitor effect against grapevine trunk diseases. Therefore, this combination of two protection strategies may constitute a promising long-term approach to mitigate the impact of Botryosphaeria dieback.

Mechanical Properties of Human Concentrated Growth Factor (CGF) Membrane and the CGF Graft with Bone Morphogenetic Protein-2 (BMP-2) onto Periosteum of the Skull of Nude Mice

Concentrated growth factor (CGF) is 100% blood-derived, cross-linked fibrin glue with platelets and growth factors. Human CGF clot is transformed into membrane by a compression device, which has been widely used clinically. However, the mechanical properties of the CGF membranes have not been well characterized. The aims of this study were to measure the tensile strength of human CGF membrane and observe its behavior as a scaffold of BMP-2 in ectopic site over the skull. The tensile test of the full length was performed at the speed of 2mm/min. The CGF membrane (5 × 5 × 2 mm3) or the CGF/BMP-2 (1.0 μg) membrane was grafted onto the skull periosteum of nude mice (5-week-old, male), and harvested at 14 days after the graft. The appearance and size of the CGF membranes were almost same for 7 days by soaking at 4 °C in saline. The average values of the tensile strength at 0 day and 7 days were 0.24 MPa and 0.26 MPa, respectively. No significant differences of both the tensile strength and the elastic modulus were found among 0, 1, 3, and 7 days. Supra-periosteal bone induction was found at 14 days in the CGF/BMP-2, while the CGF alone did not induce bone. These results demonstrated that human CGF membrane could become a short-term, sticky fibrin scaffold for BMP-2, and might be preserved as auto-membranes for wound protection after the surgery.

Evaluation of Biological and Chemical Pruning Wound Protectants to Control Grapevine Trunk Diseases Pathogens Eutypa lata and Neofusicoccum parvum

Grapevine trunk diseases, caused by many different fungal pathogens, are one of the most economically important diseases affecting the grapevine industry worldwide. Pruning wounds are the main point of entry for these fungal pathogens and thus, disease control is focused on preventative pruning wound protection by chemical products and/or biological control agents (BCAs). In this study we evaluated a broad variety of already registered or at the experimental stage of chemical and BCAs in greenhouse and in field trials for the protection of table- and wine-grape vines against infection of Eutypa lata and Neofusicoccum parvum, major pathogens responsible for Eutypa and Botryosphaeria dieback, respectively. Our study showed that Trichoderma asperellum and Trichoderma gamsii consistently provided pruning wound protection in greenhouse and field trials, with a mean percent disease control (MPDC) of 88% and 100% for E. lata and N. parvum, respectively, when compared to the water treated-inoculated positive control (P<0.05). The chemical protectants, thiophanate-methyl + myclobutanil and fluopyram and trifloxystrobin were also able to effectively protect wounds with a MPDC of up to 86% when compared to the water treated-inoculated positive control (P<0.05). When biological treatments were evaluated for recovery from treated canes at the end of the growing season, Trichoderma-based treatments had a rate of recovery between 0 and 100%, Aureobasidium-based treatments had a recovery rate between 25 and 100%, and Bacillus-based treatments had a recovery rate between 0 and 25%.

Challenges Related to Surgical Site Infection Prevention—Results after Standardized Bundle Implementation

Aim: The aim of this study was to assess the implementation of an intraoperative standardized surgical site infection (SSI) prevention bundle. Methods: The multimodal, evidence-based care bundle included nine intraoperative items (antibiotic type, timing, and re-dosing; disinfection; induction temperature control > 36.5°; glove change; intra-cavity lavage; wound protection; and closure strategy). The bundle was applied to all consecutive patients undergoing colonic resections. The primary outcome, SSI, was independently assessed by the National Infection Surveillance Committee for up to 30 postoperative days. A historical, institutional pre-implementation control group (2012–2017) with an identical methodology was used for comparison. Findings: In total, 1516 patients were included, of which 1256 (82.8%) were in the control group and 260 (17.2%) were in the post-implementation group. After 2:1 propensity score matching, the groups were similar for all items (p > 0.05). Overall compliance with the care bundle was 77% (IQR 77–88). The lowest compliance rates were observed for temperature control (53% overall), intra-cavity lavage (64% overall), and wound protection and closure (68% and 63% in the SSI group, respectively). Surgical site infections were reported in 58 patients (22.2%) vs. 21.4% in the control group (p = 0.79). Infection rates were comparable throughout the Centers for Disease Control and Prevention (CDC) categories: superficial, 12 patients (4.5%) vs. 4.2%, p = 0.82; deep incisional, 10 patients (3.7%) vs. 5.1%, p = 0.34; organ space, 36 (14%) vs. 12.4%, p = 0.48. After propensity score matching, rates remained comparable throughout all comparisons (all p > 0.05). Conclusions: The implementation of an intraoperative standardized care bundle had no impact on SSI rates. This may be explained by insufficient compliance with the individual measures.

Advance-Platelet Rich Fibrin and Hyaluronic Acid Combination Improves Interleukin-6 and Granulation Index in Diabetic Foot Ulcer Patients

BACKGROUND: Diabetic foot ulcer (DFU) is the most common and threatening complication of Diabetes Mellitus (DM). Ideal wound dressing for DFU management should relieve symptoms, provide wound protection, and encourage healing. Advanced-Platelet Rich Fibrin (A-PRF) and Hyaluronic Acid (HA) have been proven to improve wound healing process. This study was aimed to demonstrate the ability of combination of A-PRF and HA in reducing inflammation and improving DFU tissue healing.METHODS: Twenty DFU subjects were involved in this study, and divided into two groups based on the topical fibrin gel treatment: A-PRF + HA group and A-PRF only group. A-PRF was obtained by peripheral blood centrifugation. A-PRF + HA was prepared by homogening A-PRF and AH with a ratio of 1:0.6. Interleukin-6 (IL-6) level, granulation index (GI), numeric pain score (NPS), and inflammation clinical symptoms (ICS) were assessed on day-0, 3, 7 and 14.RESULTS: Wound swabs’ IL-6 level on day-7 was found to be significantly lower in A-PRF + HA compared to A-PRF alone (p=0.041). The IL-6 level reduction also found to be significant higher either in wound swabs (day 0-7, p=0.015) or fibrin gel (day 0-3, p=0.049; day 0-7, p=0.034). A-PRF + HA treatment significantly increased the GI even since day-3 (p=0.043), with lower NPS (p<0.001), and ICS score.CONCLUSION: The combination of A-PRF and HA increases the GI in DFU healing by reducing the inflammation state which will induce the angiogenesis process, as well as reducing pain in DFU subjects better than A-PRF alone.KEYWORDS: inflammation, interleukin-6, wound healing, angiogenesis, proliferation

In Vitro User Assessment of a Novel Wound Protection and Specimen Extraction System for Laparoscopic Procedures Involving Morcellation

Introduction: The risk of unknowingly disseminating leiomyosarcoma by morcellation in women undergoing laparoscopic hysterectomy has massively impacted gynaecological practice. Here, we present the results of an in vitro assessment of a novel protection system developed to mitigate this hazard. Methods: The Tissue Containment System for Manual Morcellation (Guardenia™, Advanced Surgical Concepts, Wicklow, Ireland) is an evolved wound protection/specimen extraction guarded bag system compatible with any 12mm trocar. Device use was assessed by device-naïve gynaecological and general surgeon volunteers (providing expert and inexpert morcellation cohorts, respectively) on a bench model consisting of biological tissue in a custom-built moulded rig with camera control after the operators were instructed in its use. Results: Twenty surgeons (10 gynaecologists/10 general surgeons, median duration of practice experience: 8 years, median annual number of laparoscopic operative procedures: 150 and 80, respectively) completed the user assessment. All subjects understood and correctly performed each step; i.e., (i) placement of the bag through the trocar, (ii) specimen bagging, (iii) incision extension (range 25-60 mm) after tethering the bag through the port, (iv) insertion of the device guard through the mouth of the bag after trocar removal, and (v) sufficient tissue morcellation within the bag to enable complete specimen removal (mean specimen weight 390g, range 201-1800g). There was 100% bag integrity by water-leak testing following use, despite scalpel contact with the guard in 14/20 cases (70%). Conclusion: Among first-time clinical users, this novel device enabled complete containment of morcellation debris and removal of a laparoscopic specimen, which would support further submission for regulatory approval.

Prospective surveillance after implementation of a colorectal surgical site infection prevention bundle

Objective Surgical site infections (SSI) are the most frequent complications after colorectal surgery. The aim of the present study was to evaluate the impact of a standardized SSI prevention bundle. Methods The multimodal, evidence-based care bundle included 9 intraoperative items (antibiotic type, timing and re-dosing, desinfection, induction temperature control &gt;36.5°, glove change, intracavity lavage, wound protection and closure strategy). The bundle was implemented in November 2018 and applied to all consecutive patients undergoing colonic resections. Demographics, surgical specifics and overall compliance to the care bundle were prospectively assessed until October 2020. The primary outcome SSI was defined according to the definition of the Center for Disease Control (CDC) and independently assessed by the National Infection Surveillance Committee (Swissnoso) up to 30 postoperative days. A historical, institutional pre-implementation control group (2012-2017, DOI: 10.1016/j.jhin.2018.09.011) with identical methodology was used for comparison. Results In total, 243 patients were included. The control group included 1’263 patients. Both groups were comparable regarding main demographics (age, sex, body mass index, American Society of Anaesthesiologists class) and surgical characteristics (type and duration of surgery). Overall compliance to the care bundle was 77% (IQR 77-88). Lowest compliance was observed for temperature control (48%), intracavity lavage (59%) and predefined wound closure strategy (74%). Surgical site infections were reported in 54 patients (22.2%) vs. 21.4% in the control group, p = 0.79. Infection rates were comparable throughout the CDC categories: superficial: 11 patients (4.5%) vs. 4.2%, p = 0.82, deep incisional: 9 patients (3.7%) vs. 5.1%, p = 0.34, organ space: 34 (14%) vs. 12.4%, p = 0.48. Conclusion Implementation of a standardized surgical care bundle had no impact on SSI rates according to these preliminary results. Improved compliance to individual measures may help to achieve a clinical benefit.

Propylene mesh versus acrylic resin stent for palatal wound protection following free gingival graft harvesting: a short-term pilot randomized clinical trial

Background Protection of the palatal wound is an essential step following harvesting a palatal soft tissue graft. The aim of the current pilot randomized clinical study was to assess the efficacy of using propylene mesh as protective sheet when compared to conventional custom made acrylic stent after harvesting a palatal graft. The primary outcome of this study was bleeding postoperatively and secondary outcomes were pain, healing profile of the donor site as well as patient satisfaction. Methods Between 2018 and 2019 we conducted a prospective randomized controlled trial of 24 patients with palatal defects. Two groups of 12 patients with 24 sites were included in this study and were treated with soft tissue grafting technique using free grafts harvested from the hard palate. The palatal wounds were protected with propylene mesh (test group) or custom-made acrylic palatal stent (control group). Participants were assessed for the amount and duration of bleeding, pain duration, and the risk of infection 2, 4, 6, 8, 14 days post-operatively. The trial had been registered in clinical trials.gov (NCT04348279). Results Four sites were excluded from the study as dropouts. The polypropylene mesh was more effective at reducing bleeding by (2.4 ± 1.075) and pain by (1.600 ± 0.516), while the custom-made acrylic stent reduced the bleeding (5.8 ± 1.22) and pain (7.100 ± 0.316). The decline in amount of bleeding amount (P value = 0.021) and its duration (P value = 0.001) achieved by the propylene mesh was statistically significant. There was no statistical significant difference in patient satisfaction and the duration of healing process between the 2 groups. However, the healing profile of the test group was statistically significant when compared with the control group (P value = 0.002). Conclusions Propylene mesh is a promising material for protection of the palatal wound due to its light weight, limited bacterial wicking, tissue compatibility. Further studies are required to adequally assess the benefits of this material in periodontal plastic surgeries.

In vitro evaluation of grapevine endophytes, epiphytes and sap micro-organisms for potential use to control grapevine trunk disease pathogens

SummaryGrapevine trunk diseases (GTDs) threaten the economic sustainability of viticulture worldwide causing a significant reduction of both yields and quality of grapes. Biological control presents a promising sustainable alternative to cultural and chemical methods to mitigate the effects of pathogens causing GTDs, including Botryosphaeria dieback, Eutypa dieback and Esca. This study aimed to identify naturally occurring potential biological control agents from a variety of grapevine tissues, including sap, cane and pith and evaluate their antagonistic activity against selected fungal pathogens responsible for GTDs in vitro. Bacterial and fungal isolates were preliminary screened in vitro to determine their antifungal activity via a dual culture assay against Neofusicoccum parvum and Eutypa lata. Among the fungal isolates, Trichoderma spp. inhibited E. lata mycelial growth up to 64% and N. parvum mycelial growth up to 73% with overgrowth and stopped growth being the likely antagonistic mechanisms. Among the bacterial isolates, Bacillus spp. inhibited E. lata mycelial growth up to 20% and N. parvum mycelial growth up to 40%. Select antagonistic isolates of Trichoderma, Bacillus and Aureobasidium spp. were subject to further dual culture antifungal analysis against Diplodia seriata and Diaporthe ampelina, with Trichoderma isolates consistently causing the greatest inhibition. Volatile organic compound antifungal analysis revealed that these Trichoderma isolates resulted significantly inhibited mycelial growth of N. parvum, E. lata and D. ampelina causing up to 20.11%, 60.55% and 70.9% inhibition respectively (P≤0.05). Multilocus sequence analysis revealed that the Trichoderma isolates are most closely related to Trichoderma asperellum and Trichoderma hamatum. This study identifies grapevine sap as a novel source of potential biological control agents for control of GTDs to support existing efforts to control GTDs. Further testing will be necessary to fully characterize these microbes mode of antagonism and assess their efficacy for pruning wound protection in planta.