Lyophilised Platelet-Rich Fibrin: Physical and Biological Characterisation

Background: Platelet-rich fibrin (PRF) has gained popularity in craniofacial surgery, as it provides an excellent reservoir of autologous growth factors (GFs) that are essential for bone regeneration. However, the low elastic modulus, short-term clinical application, poor storage potential and limitations in emergency therapy use restrict its more widespread clinical application. This study fabricates lyophilised PRF (Ly-PRF), evaluates its physical and biological properties, and explores its application for craniofacial tissue engineering purposes. Material and methods: A lyophilisation method was applied, and the outcome was evaluated and compared with traditionally prepared PRF. We investigated how lyophilisation affected PRF’s physical characteristics and biological properties by determining: (1) the physical and morphological architecture of Ly-PRF using SEM, and (2) the kinetic release of PDGF-AB using ELISA. Results: Ly-PRF exhibited a dense and homogeneous interconnected 3D fibrin network. Moreover, clusters of morphologically consistent cells of platelets and leukocytes were apparent within Ly-PRF, along with evidence of PDGF-AB release in accordance with previously reports. Conclusions: The protocol established in this study for Ly-PRF preparation demonstrated versatility, and provides a biomaterial with growth factor release for potential use as a craniofacial bioscaffold.     

Multi-Material Scaffolds for Tissue Engineering

A trend in developing biocompatible scaffolds for tissue engineering has been to seek an ideal single material for which a given cell type will exhibit favorable behavior. While an ideal single material has proven elusive, scaffold manufacture using combinations of specialist materials can produce more versatile structures. By controlling the percentage and architecture of material components, mechanical properties, cell attachment, and proliferation may be optimized for a given function. Three specialist materials, poly-ϵ-caprolactone (PCL), fibrin, and alginate, were incorporated into multi-component scaffolds for a series of experiments testing each component with culture of fibroblasts. The rigid and formable PCL provided structure, the fibrin pore-filler allowed for cell attachment, and alginate thread provided a nutrient transfer pathway in lieu of a vascular system. The efficacy of these scaffolds was judged on fibroblast distribution and population after 7-12 days of culture.     

Injectable Hydrogels for Cardiac Tissue Engineering

In light of the limited efficacy of current treatments for cardiac regeneration, tissue engineering approaches have been explored for their potential to provide mechanical support to injured cardiac tissues, deliver cardio‐protective molecules, and improve cell‐based therapeutic techniques. Injectable hydrogels are a particularly appealing system as they hold promise as a minimally invasive therapeutic approach. Moreover, injectable acellular alginate‐based hydrogels have been tested clinically in patients with myocardial infarction (MI) and show preservation of the left ventricular (LV) indices and left ventricular ejection fraction (LVEF). This review provides an overview of recent developments that have occurred in the design and engineering of various injectable hydrogel systems for cardiac tissue engineering efforts, including a comparison of natural versus synthetic systems with emphasis on the ideal characteristics for biomimetic cardiac materials.     

Chitosan Based Scaffolds by Lyophilization and sc.CO2 Assisted Methods for Tissue Engineering Applications

Porous chitosan scaffolds with possible tissue engineering applications were synthesized by using lyophilization and supercritical carbon dioxide (sc.CO₂) drying technique. 1% Chitosan (CS) solution in aq. acetic acid was treated with 37% formaldehyde solution; the resulting hydrogels were subjected to solvent-exchange prior to the final treatment procedures. Their morphology, pore structure, and physical properties were characterized by Fourier transform infrared spectroscopy (FTIR), thermal analysis, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and the specific surface areas and porosities of scaffolds were determined by using N₂ adsorption. The sc.CO₂ treated scaffolds showed a much greater surface area in comparison to the lyophilized one. Hence, sc.CO₂ treated scaffolds is better for cell proliferation. We further investigated the bioactivity of sc.CO₂ treated scaffolds using simulated body fluid (SBF). The sc.CO₂ assisted chitosan scaffold prepared by using green chemistry approach is highly pure and from a hygienic point of view, it is an ideal material for tissue engineering applications.     

Recent Advances in Biomaterials-Based Therapies for Alleviation and Regeneration of Traumatic Brain Injury

Traumatic brain injury (TBI), a major public health problem accompanied with numerous complications, usually leads to serve disability and huge financial burden. The adverse and unfavorable pathological environment triggers a series of secondary injuries, resulting in serious loss of nerve function and huge obstacle of endogenous nerve regeneration. With the advances in adaptive tissue regeneration biomaterials, regulation of detrimental microenvironment to reduce the secondary injury and to promote the neurogenesis becomes possible. The adaptive biomaterials could respond and regulate biochemical, cellular, and physiological events in the secondary injury, including excitotoxicity, oxidative stress, and neuroinflammation, to rebuild circumstances suitable for regeneration. In this review, the development of pathology after TBI is discussed, followed by the introduction of adaptive biomaterials based on various pathological characteristics. The adaptive biomaterials carried with neurotrophic factors and stem cells for TBI treatment are then summarized. Finally, the current drawbacks and future perspective of biomaterials for TBI treatment are suggested.     

Advances in Mechanical Properties of Hydrogels for Cartilage Tissue Defect Repair

Numerous factors, such as degeneration and accidents, frequently cause cartilage deterioration. Owing to the absence of blood vessels and nerves in cartilage tissue, the ability of cartilage tissue to heal itself after an injury is relatively low. Hydrogels are beneficial for cartilage tissue engineering owing to their cartilage-like structure and advantageous properties. Due to the disruption of its mechanical structure, the bearing capacity and shock absorption of cartilage are diminished. The tissue should possess excellent mechanical properties to ensure the efficacy of cartilage tissue repair. This paper discusses the application of hydrogels in the fields of cartilage repair, the mechanical properties of hydrogels used for cartilage repair, and the materials used for hydrogels in cartilage tissue engineering. In addition, the challenges faced by hydrogels and future research directions are discussed.     

Development of Channeled Nanofibrous Scaffolds for Oriented Tissue Engineering

A tissue‐engineering scaffold resembling the structure of the natural extracellular matrix can often facilitate tissue regeneration. Nerve and tendon are oriented micro‐scale tissue bundles. In this study, a method combining injection molding and thermally induced phase separation techniques is developed to create single‐ and multiple‐channeled nanofibrous poly(L ‐lactic acid) scaffolds. The overall shape, the number and spatial arrangement of channels, the channel wall matrix architecture, the porosity and mechanical properties of the scaffolds are all tunable. The porous NF channel wall matrix provides an excellent microenvironment for protein adsorption and the attachment of PC12 neuronal cells and tendon fibroblast cells, showing potential for neural and tendon tissue regeneration.

     

Polypyrrole/Alginate Hybrid Hydrogels: Electrically Conductive and Soft Biomaterials for Human Mesenchymal Stem Cell Culture and Potential Neural Tissue Engineering Applications

Electrically conductive biomaterials that can efficiently deliver electrical signals to cells or improve electrical communication among cells have received considerable attention for potential tissue engineering applications. Conductive hydrogels are desirable particularly for neural applications, as they can provide electrical signals and soft microenvironments that can mimic native nerve tissues. In this study, conductive and soft polypyrrole/alginate (PPy/Alg) hydrogels are developed by chemically polymerizing PPy within ionically cross‐linked alginate hydrogel networks. The synthesized hydrogels exhibit a Young's modulus of 20–200 kPa. Electrical conductance of the PPy/Alg hydrogels could be enhanced by more than one order of magnitude compared to that of pristine alginate hydrogels. In vitro studies with human bone marrow‐derived mesenchymal stem cells (hMSCs) reveal that cell adhesion and growth are promoted on the PPy/Alg hydrogels. Additionally, the PPy/Alg hydrogels support and greatly enhance the expression of neural differentiation markers (i.e., Tuj1 and MAP2) of hMSCs compared to tissue culture plate controls. Subcutaneous implantation of the hydrogels for eight weeks induces mild inflammatory reactions. These soft and conductive hydrogels will serve as a useful platform to study the effects of electrical and mechanical signals on stem cells and/or neural cells and to develop multifunctional neural tissue engineering scaffolds.

     

Design of a Bioabsorbable Multilayered Patch for Esophagus Tissue Engineering

A gold standard for esophagus reconstruction is not still available. The present work aims to design a polymer patch combining synthetic polylactide‐co‐polycaprolacton and chitosan biopolymers, tailoring patch properties to esophageal tissue characteristics by a temperature‐induced precipitation method, to get multilayered patches (1L, 2L, and 3L). Characterization shows stable multilayered patches (1L and 2L) by selection of copolymer type, and their M _w. In vitro investigation of the functional patch properties in simulated physiologic and pathologic conditions demonstrates that the chitosan layer (patch 3L) decreases patch stability and cell adhesion, while improves cell proliferation. Patches 2L and 3L comply with physiological esophageal pressure (3–5 kPa) and elongation (20%).

     

Development of an Injectable Calcium Phosphate/Hyaluronic Acid Microparticles System for Platelet Lysate Sustained Delivery Aiming Bone Regeneration

Despite the biocompatibility and osteoinductive properties of calcium phosphate (CaP) cements their low biodegradability hampers full bone regeneration. Herein the incorporation of CaP cement with hyaluronic acid (HAc) microparticles loaded with platelet lysate (PL) to improve the degradability and biological performance of the cements is proposed. Cement formulations incorporating increasing weight ratios of either empty HAc microparticles or microparticles loaded with PL (10 and 20 wt%) are developed as well as cements directly incorporating PL. The direct incorporation of PL improves the mechanical properties of the plain cement, reaching values similar to native bone. Morphological analysis shows homogeneous particle distribution and high interconnectivity between the HAc microparticles. The cements incorporating PL (with or without the HAc microparticles) present a sustained release of PL proteins for up to 8 d. The sustained release of PL modulates the expression of osteogenic markers in seeded human adipose tissue derived stem cells, thus suggesting the stimulatory role of this hybrid system toward osteogenic commitment and bone regeneration applications.

     

Evaluation of PEGylated fibrin as a three-dimensional biodegradable scaffold for ovarian tissue engineering

The most challenging task of creating a bioengineered ovary to restore fertility in cancer patients is choosing an appropriate biomaterial to encapsulate isolated preantral follicles and ovarian cells. In this study, as a biocompatible and biodegradable biomaterial containing fibrin-like bioactivity and manageable physical properties, PEGylated fibrin aims to encapsulate isolated ovarian stromal cells as a first step of creating an engineered ovarian tissue. For this purpose, human ovarian stromal cells were isolated from frozen-thawed ovarian tissue and cultured in the PEGylated fibrin hydrogels (PEG:Fib), which were fabricated by combining two different molar ratios of PEG:Fib (10:1 and 5:1) and two thrombin concentrations. The samples were analyzed at days 0 and 5 of in vitro for cell density, proliferation (Ki67), and apoptosis (caspase-3). Moreover, LIVE/DEAD and PrestoBlue assays assessed cell viability and proliferation on days 1, 3, and 5. The effect of PEGylation on the biodegradation behavior of fibrin was evaluated by measuring the remaining mass ratio of non-modified fibrin, PEG:Fib 10:1, and PEG:Fib 5:1 hydrogels after 1, 2, 3, 5, 8, 11, and 15 days. The results showed that PEGylated fibrin hydrogels enhanced scaffold stability and supported cell viability and proliferation. In addition, PEG:Fib 5:1 T50 indicated a significantly higher cell density dynamic and non-significantly lower expression of caspase-3 on day 5. Besides, uniformity of cell distribution inside the hydrogel and a tendency to a high rate of Ki67-positive cells was observed in PEG:Fib 10:1 T50 hydrogels. In conclusion, this study reveals the positive effects of PEGylated fibrin hydrogels on isolated human ovarian stromal cells. Based on such promising findings, we believe that this matrix should be tested to encapsulate isolated human ovarian follicles.     

Functionalization of Biodegradable Polyester for Tissue Engineering Applications

Summary: Chemical modification of polymer surface may potentially be used to create smart materials that can guide cellular adhesion, proliferation and maintenance of specific expression of molecules. The microbial polyester poly (3-hydroxybutyrate) (PHB) has been attracted attention as promising material for applications in tissue engineering. In this work, a wet-chemical method, base ethylenediamine aminolysis, was performed to improve the adhesion of chondrocytes isolated from human articular cartilage to PHB films. The effects of chemical treatment on PHB films was evaluated by following changes in morphology and surface chemical composition using atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS), respectively. While the effect on cells morphology was studied by scanning electron microscopy (SEM). The treatment with ethylenediamine did not change significantly the morphology of the structures of PHB films surface. However, the roughness of the aminolyzed films was slightly higher. The introduction of nitrogen-containing groups was confirmed by XPS. In vitro experiments indicated that the surface modification did not have toxic effects in cells, since they could adhere and proliferate on modified PHB films. It was observed that long-time treatment improved ability of PHB films to support cell growth, which could be accounted to physicochemical and topological effects.     

Electroactive Composite of FeCl3‐Doped P3HT/PLGA with Adjustable Electrical Conductivity for Potential Application in Neural Tissue Engineering

Conducting polymers (CPs) is one of intelligent biomaterials with the specific properties of reversible redox states, which have a significant effects on the cell behaviors and nerve tissue regeneration. However, the effects of CPs with different electrical conductivity on the behaviors of nerve cells are rarely reported. Therefore, a kind of Poly(3‐hexylthiophene) (P3HT) with certain molecular weight is synthesized by Kumada catalyst transfer polymerization (KCTP) method and employed to prepare bioabsorbable and electroactive intelligent composites of Poly(3‐hexylthiophene)/Poly(glycolide‐lactide) (P3HT/PLGA). FeCl₃ doping electroactive membranes with different electrical conductivities are prepared to investigate the cell behaviors. On the substrate with higher electrical conductivity, the proliferation of rat adrenal pheochromocytoma cells (PC12 cells) is significantly promoted and neurite length is increased obviously. In particular, the most significant improvements are the neuron gene expression of Synapsin 1 and microtubule‐associated protein 2 (MAP2) by the composites with high conductivity. These results suggest that P3HT/PLGA with suitable electrical conductivity have a positive role in promoting neural growth and differentiation, which is promising for advancing potential application of nerve repair and regeneration.     

Development of 3D-Printed,Biodegradable, Conductive PGSA Composites for Nerve Tissue Regeneration

Nerve conduits are used to reconnect broken nerve bundles and provide protection to facilitate nerve regeneration. However, the low degradation rate and regeneration rate, as well as the requirement for secondary surgery are some of the most criticized drawbacks of existing nerve conduits. With high processing flexibility from the photo-curability, poly (glycerol sebacate) acrylate (PGSA) is a promising material with tunable mechanical properties and biocompatibility for the development of medical devices. Here, polyvinylpyrrolidone (PVP), silver nanoparticles (AgNPs), and graphene are embedded in biodegradable PGSA matrix. The polymer composites are then assessed for their electrical conductivity, biodegradability, three-dimensional-printability (3D-printability), and promotion of cell proliferation. Through the four-probe technique, it is shown that the PGSA composites are identified as highly conductive in swollen state. Furthermore, biodegradability is evaluated through enzymatic degradation and facilitated hydrolysis. Cell proliferation and guidance are significantly promoted by three-dimensional-printed microstructures and electrical stimulation on PGSA composites, especially on PGSA-PVP. Hence, microstructured nerve conduits are 3D-printed with PGSA-PVP. Guided cell growth and promoted proliferation are subsequently demonstrated by Schwann cell culture combined with electrical stimulation. Consequently, 3D-printed nerve conduits fabricated with PGSA composites hold great potential in nerve tissue regeneration through electrical stimulation.     

组织工程相关生物材料表面工程的研究进展

生物材料用作人工细胞外基质(ECM ) 在组织工程中占据重要位置。本文在分析细胞2生物材料表面相互作用的基础上, 从生物材料中的水、材料表面的形态、材料表面的特异性识别及生物材料诱发愈合等方面探讨了生物材料的复杂性。生物材料对细胞的影响是一个双向、动态过程, 起着调节细胞增殖和凋亡平衡的作用。基于生物材料对细胞生长的影响, 本文提出了生物材料表面生物仿生化以提高细胞亲和力,糖链团簇、糖脂质及材料表面蛋白质修饰以提高细胞特异性识别, 材料表面的自组装修饰以改善表面形态等观点。     

Influence of Materials Properties on Bio-Physical Features and Effectiveness of 3D-Scaffolds for Periodontal Regeneration

Periodontal diseases are multifactorial disorders, mainly due to severe infections and inflammation which affect the tissues (i.e., gum and dental bone) that support and surround the teeth. These pathologies are characterized by bleeding gums, pain, bad breath and, in more severe forms, can lead to the detachment of gum from teeth, causing their loss. To date it is estimated that severe periodontal diseases affect around 10% of the population worldwide thus making necessary the development of effective treatments able to both reduce the infections and inflammation in injured sites and improve the regeneration of damaged tissues. In this scenario, the use of 3D scaffolds can play a pivotal role by providing an effective platform for drugs, nanosystems, growth factors, stem cells, etc., improving the effectiveness of therapies and reducing their systemic side effects. The aim of this review is to describe the recent progress in periodontal regeneration, highlighting the influence of materials’ properties used to realize three-dimensional (3D)-scaffolds, their bio-physical characteristics and their ability to provide a biocompatible platform able to embed nanosystems.     

Large-Scale Production of 3D Bioactive Glass Macroporous Scaffolds for Tissue Engineering

For tissue engineering applications, a scaffold is required that can act as a template and guide for cell proliferation, cell differentiation and tissue growth. Interconnected pores with diameters greater than 100 m are required for tissue ingrowth, vascularisation and nutrient delivery to the centre of the scaffold. 3D bioactive glass scaffolds have been produced, by foaming sol-gel derived bioactive glasses. The method to produce foams with a modal macropore diameter of 100 m, and a handling strength suitable for cell culture, was to foam 50 ml batches of sol with the aid of a surfactant and gelling agent. In vitro and in vivo tests show that the scaffolds have high potential to be used in bone tissue engineering applications. Larger batches are required to produce scaffolds commercially. The aim of this work was to investigate how the process could be up-scaled for commercial use. This study shows that foaming larger aliquots of sol decreased the scaffold porosity and interconnectivity and investigates methods of modifying the process to obtain large quantities of foam scaffolds with pores in excess of 100 m. The optimum method to produce foams of similar pore structure from 200 ml sol to those produced from 50 ml sol comprised of adding 3 ml surfactant and 12 ml dionised water to the sol to start foaming and injecting a gas mixture (70% helium, 30% nitrogen) at 0.2 bar while applying vigorous agitation.     

Three-Dimensional Bioprinting of Decellularized Extracellular Matrix-Based Bioinks for Tissue Engineering

Three-dimensional (3D) bioprinting is one of the most promising additive manufacturing technologies for fabricating various biomimetic architectures of tissues and organs. In this context, the bioink, a critical element for biofabrication, is a mixture of biomaterials and living cells used in 3D printing to create cell-laden structures. Recently, decellularized extracellular matrix (dECM)-based bioinks derived from natural tissues have garnered enormous attention from researchers due to their unique and complex biochemical properties. This review initially presents the details of the natural ECM and its role in cell growth and metabolism. Further, we briefly emphasize the commonly used decellularization treatment procedures and subsequent evaluations for the quality control of the dECM. In addition, we summarize some of the common bioink preparation strategies, the 3D bioprinting approaches, and the applicability of 3D-printed dECM bioinks to tissue engineering. Finally, we present some of the challenges in this field and the prospects for future development.