Evaluation of the Biocompatibility of Cleaned Bovine Tendon in a Heterotopic Implantation Model

Damage to the tendon-ligamentous apparatus places serious limitations on a personʼs physical activity. Injuries are especially common in physically healthy people leading an active lifestyle, such as athletes. To treat such injuries in orthopaedics and traumatology, autoplastic operations are performed or prostheses made of synthetic or biological materials are installed. The known treatment methods, in spite of their effectiveness, have a number of serious drawbacks, which often limit their use. Therefore, the search for new approaches and materials for plastic ligaments is an urgent task. Today, biotissue prostheses are accumulating advantages over their synthetic counterparts. The most promising raw material for biological ligament prostheses, due to its availability in the required quantity and optimal size, is the flexor and extensor calf tendons. This paper aimed to develop a method for treating xenogenic tendon to manufacture ligament prostheses and assessing its biocompatibility in a heterotopic implantation model. To manufacture a ligament prosthesis, the raw material was subjected to mechanical cleaning and chemical-physical treatment, as well as treatment with supercritical carbon dioxide fluid with the addition of the nonionic surfactant Tween 80, which together contributed to effective decellularization and removal of other biologically active components, while maintaining the physical and mechanical parameters and natural fiberarchitectonics of native raw materials. The biocompatible properties of ligament prosthesis specimens made from the flexor and extensor calf tendons using this method were evaluated in a model of heterotopic implantation into the subcutaneous adipose tissue of rats. The results obtained confirm the promising use of this material, treated according to the proposed method, in clinical practice.

Elucidating mechanisms of antitumor immunity mediated by live oncolytic vaccinia and heat-inactivated vaccinia

BackgroundViral-based immunotherapy can overcome resistance to immune checkpoint blockade (ICB) and fill the unmet needs of many patients with cancer. Oncolytic viruses (OVs) are defined as engineered or naturally occurring viruses that selectively replicate in and kill cancer cells. OVs also induce antitumor immunity. The purpose of this study was to compare the antitumor effects of live oncolytic vaccinia viruses versus the inactivated versions and elucidate their underlying immunological mechanisms.MethodsWe engineered a replication-competent, oncolytic vaccinia virus (OV-GM) by inserting a murine GM-CSF gene into the thymidine kinase locus of a mutant vaccinia E3L∆83N, which lacks the Z-DNA-binding domain of vaccinia virulence factor E3. We compared the antitumor effects of intratumoral (IT) delivery of live OV-GM versus heat-inactivated OV-GM (heat-iOV-GM) in a murine B16-F10 melanoma bilateral implantation model. We also generated vvDD, a well-studied oncolytic vaccinia virus, and compared the antitumor effects of live vvDD vs heat-inactivated vvDD (heat-ivvDD) in a murine A20 B-cell lymphoma bilateral tumor implantation model.ResultsHeat-iOV-GM infection of dendritic cells (DCs) and tumor cells in vitro induced type I interferon and proinflammatory cytokines and chemokines, whereas live OV-GM did not. IT live OV-GM was less effective in generating systemic antitumor immunity compared with heat-iOV-GM. Similar to heat-iOV-GM, the antitumor effects of live OV-GM also require Batf3-dependent CD103+ dendritic cells. When combined with systemic delivery of ICB, IT heat-iOV-GM was more effective in eradicating tumors, compared with live OV-GM. IT heat-ivvDD was also more effective in treating murine A20 B-cell lymphoma, compared with live vvDD.ConclusionsTumor lysis induced by the replication of oncolytic vaccinia virus has a limited effect on the generation of systemic antitumor immunity. The activation of Batf3-dependent CD103+ DCs is critical for antitumor effects induced by both live OV-GM and heat-iOV-GM, with the latter being more potent than live OV-GM in inducing innate and adaptive immunity in both locally injected and distant, non-injected tumors. We propose that evaluations of both innate and adaptive immunity, induced by IT oncolytic viral immunotherapy at injected and non-injected tumors, should be included as potential biomarkers for host responses to viral therapy.

Ex Vivo and In Vivo Analysis of a Novel Porcine Aortic Patch for Vascular Reconstruction

(1) Background: The aim of the present study was the biocompatibility analysis of a novel xenogeneic vascular graft material (PAP) based on native collagen won from porcine aorta using the subcutaneous implantation model up to 120 days post implantationem. As a control, an already commercially available collagen-based vessel graft (XenoSure®) based on bovine pericardium was used. Another focus was to analyze the (ultra-) structure and the purification effort. (2) Methods: Established methodologies such as the histological material analysis and the conduct of the subcutaneous implantation model in Wistar rats were applied. Moreover, established methods combining histological, immunohistochemical, and histomorphometrical procedures were applied to analyze the tissue reactions to the vessel graft materials, including the induction of pro- and anti-inflammatory macrophages to test the immune response. (3) Results: The results showed that the PAP implants induced a special cellular infiltration and host tissue integration based on its three different parts based on the different layers of the donor tissue. Thereby, these material parts induced a vascularization pattern that branches to all parts of the graft and altogether a balanced immune tissue reaction in contrast to the control material. (4) Conclusions: PAP implants seemed to be advantageous in many aspects: (i) cellular infiltration and host tissue integration, (ii) vascularization pattern that branches to all parts of the graft, and (iii) balanced immune tissue reaction that can result in less scar tissue and enhanced integrative healing patterns. Moreover, the unique trans-implant vascularization can provide unprecedented anti-infection properties that can avoid material-related bacterial infections.

In Vivo Tissue Implantation Model of In Vivo Tissue Implantation Model of Lidocaine (LCH)-Encapsulated Dextran (DEX)/Glycol (GLY) Nanoparticles Delivery for Pain Management

Lidocaine-loaded nanoparticles are versatile nanomaterials that may be used in pain treatment due to their wound healing properties. The current study describes a wound dressing formulation focused on lidocaine-loaded dextran/ethylene glycol nanoparticles (an anesthetic drug). The lidocaine-loaded dextran/ethylene glycol membranes were fabricated using lidocaine solutions inside the dextran/ethylene glycol medium. The influence of various experimental conditions on dextran/ethylene glycol nanoparticle formations were examined. The sizes of dextran/ethylene glycol and lidocaine-loaded dextran/glycol nanoparticles were examined through the HR-SEM. Moreover, the efficacy antibacterial activity of dextran/glycol and lidocaine-loaded dextran/ethylene glycol nanoparticles was evaluated against the microorganisms grampositive and negative. Furthermore, we observed the In Vivo wound healing of wounds in skin using a mice model over a 16 days period. In this difference to the wounds of untreated mouse, quick healing was observed in the lidocaine-loaded dextran/glycol nanoparticles-treated wounds with fewer injury. These results specify that lidocaine-loaded dextran/ethylene glycol nanoparticles-based dressing material could be a ground-breaking nanomaterial having wound repair and implantations potential required for wound injury in pain management, which was proven using an animal model.

Ex Vivo and In Vivo Analyses of Novel 3D-Printed Bone Substitute Scaffolds Incorporating Biphasic Calcium Phosphate Granules for Bone Regeneration

(1) Background: The aim of this study was examining the ex vivo and in vivo properties of a composite made from polycaprolactone (PCL) and biphasic calcium phosphate (BCP) (synprint, ScientiFY GmbH) fabricated via fused deposition modelling (FDM); (2) Methods: Scaffolds were tested ex vivo for their mechanical properties using porous and solid designs. Subcutaneous implantation model analyzed the biocompatibility of PCL + BCP and PCL scaffolds. Calvaria implantation model analyzed the osteoconductive properties of PCL and PCL + BCP scaffolds compared to BCP as control group. Established histological, histopathological and histomorphometrical methods were performed to evaluate new bone formation.; (3) Results Mechanical testing demonstrated no significant differences between PCL and PCL + BCP for both designs. Similar biocompatibility was observed subcutaneously for PCL and PCL + BCP scaffolds. In the calvaria model, new bone formation was observed for all groups with largest new bone formation in the BCP group, followed by the PCL + BCP group, and the PCL group. This finding was influenced by the initial volume of biomaterial implanted and remaining volume after 90 days. All materials showed osteoconductive properties and PCL + BCP tailored the tissue responses towards higher cellular biodegradability. Moreover, this material combination led to a reduced swelling in PCL + BCP; (4) Conclusions: Altogether, the results show that the newly developed composite is biocompatible and leads to successful osteoconductive bone regeneration. The new biomaterial combines the structural stability provided by PCL with bioactive characteristics of BCP-based BSM. 3D-printed BSM provides an integration behavior in accordance with the concept of guided bone regeneration (GBR) by directing new bone growth for proper function and restoration.