Chitosan composite scaffolds for articular cartilage defect repair: a review

Articular cartilage (AC) defects lack the ability to self-repair due to their avascular nature and the declined mitotic ability of mature chondrocytes. To date, cartilage tissue engineering using implanted scaffolds containing cells or growth factors is the most promising defect repair method. Scaffolds for cartilage tissue engineering have been comprehensively researched. As a promising scaffold biomaterial for AC defect repair, the properties of chitosan are summarized in this review. Strategies to composite chitosan with other materials, such as polymers (including collagen, gelatin, alginate, silk fibroin, poly-caprolactone, and poly-lactic acid) and bioceramics (including calcium phosphate, calcium polyphosphate, and hydroxyapatite) are presented. Methods to manufacture three-dimensional porous structures to support cell attachment and nutriment exchange have also been included.

Properties of chitosan/polymer and chitosan/bioceramic composite scaffolds for articular cartilage defect repair are reviewed.     

Synergistic delivery of bFGF and BMP-2 from poly(l-lactic-co-glycolic acid)/graphene oxide/hydroxyapatite nanofibre scaffolds for bone tissue engineering applications

One of the goals of bone tissue engineering is to create scaffolds with excellent biocompatibility, osteoinductive ability and mechanical properties. The application of bioactive proteins, such as bone morphogenetic protein (BMP)-2 and basic fibroblast growth factor (bFGF), has been showed to be an effective way to improve the osteoinductivity and biocompatibility of bone scaffold materials. Therefore, the development of novel materials capable of delivering multiple growth factors is urgent and essential for bone defect repair. In this study, a composite nanofibre scaffold composed of poly(l-lactic-co-glycolic acid) (PLGA), hydroxyapatite (HA), and graphene oxide (GO) has been fabricated to deliver basic fibroblast growth factor (bFGF) and bone morphogenetic protein-2 (BMP-2) simultaneously. The data show that the incorporation of GO and HA into PLGA nanofibres significantly improved the mechanical properties and hydrophilicity of the nanofibre scaffolds. More importantly, compared to PLGA and PLGA/HA nanofibre scaffolds, the PLGA/HA/GO nanofibre scaffolds could more efficiently immobilize bFGF and BMP-2. Moreover, biological assays indicated that the loaded bFGF and BMP-2 loaded in the composite nanofibre scaffolds have a synergistic differentiation effect on the cell adhesion, proliferation, and osteogenesis differentiation of MC3T3-E1 cells. In contrast to the PLGA/HA/GO/bFGF and PLGA/HA/GO/BMP-2 nanofibre scaffolds, the PLGA/HA/GO/bFGF/BMP-2 scaffolds have shown higher ALP activity and higher expression levels of osteogenesis-related genes. In summary, our findings indicated that the incorporation of GO into nanofibre scaffolds is an effective method to immobilize growth factors onto biomaterial surfaces, and the synergistic effects of a combination of BMP-2 and bFGF may have potential use in bone regenerative therapeutics.

One of the goals of bone tissue engineering is to create scaffolds with excellent biocompatibility, osteoinductive ability and mechanical properties.     

Preparation and characterization of a novel drug-loaded Bi-layer scaffold for cartilage regeneration

The incidence of articular cartilage defects is increasing year by year. In order to repair the cartilage tissue at the defect, scaffolds with nanofiber structure and biocompatibility have become a research hotspot. In this study, we designed and fabricated a bi-layer scaffold prepared from an upper layer of drug-dispersed gelatin methacrylate (GELMA) hydrogel and a lower layer of a drug-encapsulated coaxial fiber scaffold prepared from silk fiber (SF) and polylactic acid (PLA). These bi-layer scaffolds have porosity (91.26 ± 3.94%) sufficient to support material exchange and pore size suitable for cell culture and infiltration, as well as mechanical properties (2.65 ± 0.31 MPa) that meet the requirements of cartilage tissue engineering. The coaxial fiber structure exhibited excellent drug release properties, maintaining drug release for 14 days in PBS. In vitro experiments indicated that the scaffolds were not toxic to cells and were amenable to chondrocyte migration. Notably, the growth of cells in a bi-layer scaffold presented two states. In the hydrogel layer, cells grow through interconnected pores and take on a connective tissue-like shape. In the coaxial fiber layer, cells grow on the surface of the coaxial fiber mats and appeared tablet-like. This is similar to the structure of the functional partitions of natural cartilage tissue. Together, the bi-layer scaffold can play a positive role in cartilage regeneration, which could be a potential therapeutic choice to solve the current problems of clinical cartilage repair.

The picture shows the preparation process of the cartilage scaffold, in which the hydrogel layer is loaded with aescin sodium, and the fiber layer is loaded with kartogenin.     

Fabrication and functionalization of 3D-printed soft and hard scaffolds with growth factors for enhanced bioactivity

Strategies to improve the acceptance of scaffolds by the body is crucial in tissue engineering (TE) which requires tailoring of the pore structure, mechanical properties and surface characteristics of the scaffolds. In the current study we used a 3-dimensional (3D) printing technique to tailor the pore structure and mechanical properties of (i) nanocellulose based hydrogel scaffolds for soft tissue engineering and (ii) poly lactic acid (PLA) based scaffolds for hard tissue engineering in combination with surface treatment by protein conjugation for tuning the scaffold bioactivity. Dopamine coating of the scaffolds enhanced the hydrophilicity and their capability to bind bioactive molecules such as fibroblast growth factor (FGF-18) for soft TE scaffolds and arginyl glycyl aspartic acid (RGD) peptide for hard TE scaffolds, which was confirmed using MALDI-TOFs. This functionalization approach enhanced the performance of the scaffolds and provided antimicrobial activity indicating that these scaffolds can be used for cartilage or bone regeneration applications. Blood compatibility studies revealed that both the materials were compatible with human red blood cells. Significant enhancement of cell attachment and proliferation confirmed the bioactivity of growth factor functionalized 3D printed soft and hard tissues. This approach of combining 3D printing with biological tuning of the interface is expected to significantly advance the development of biomedical materials related to soft and hard tissue engineering.

3D printed scaffolds with tailored bioactivity using protein conjugation.     

Three-dimensional cell printing of gingival fibroblast/acellular dermal matrix/gelatin–sodium alginate scaffolds and their biocompatibility evaluation in vitro

Tissue engineering has emerged as a promising approach for soft tissue regeneration. Three-dimensional (3D) cell printing showed great potential for producing cell-encapsulated scaffolds to repair tissue defects. The advantage of 3D cell printing technology is precise cell loading in scaffolds to achieve tissue regeneration instead of only relying on the cells from surrounding tissue or blood. A new acellular dermal matrix/gelatin–sodium alginate (ADM/A/G) scaffold with living gingival fibroblasts was constructed by 3D cell printing technology for potential oral soft tissue regeneration in this study, and the biological characteristics of the 3D cell printing scaffolds were evaluated. The residue of nucleic acid and growth factors in ADM were detected. Three biomaterials were mixed at an appropriate radio with human gingival fibroblasts (hGFs) to prepare bioinks. Two kinds of layer scaffolds were fabricated by 3D cell printing technology. The mechanical strength and degradability of the scaffolds were determined by measuring their compressive modulus and mass loss. CCK-8 assay and calcein-AM/PI staining were conducted to detect the cell proliferation and viability in 3D cell printing scaffolds. The morphology of the hGFs in the scaffolds were observed using SEM and FITC-phalloidin staining. The expression of COL1A1, PECAM1, and VEGF-A of hGFs in the scaffolds were quantified by qRT-PCR. The gelatin–sodium alginate (A/G) scaffolds were used as control group in all experiments. Compared with the control group, 3D cell printing ADM/A/G scaffolds showed better mechanical strength and longer degradation time. The ADM/A/G scaffolds obviously had a better promotion effect on cell proliferation and viability. Most of the hGFs observed had a fully extended spindle morphology in the ADM/A/G scaffolds but oval morphology in the control group. The expression of COL1A1 was significantly higher than in the control group with time, and the expression of PECAM1 and VEGF-A was slightly higher in ADM/A/G scaffolds on day 14. 3D cell printing gingival fibroblast-ADM/A/G scaffolds showed excellent biological properties, which could be potentially useful in oral soft tissue regeneration.

3D cell printing of gingival fibroblast/acellular dermal matrix/gelatin–sodium alginate scaffolds showed satisfactory biological properties.     

Polydopamine-modified poly(l-lactic acid) nanofiber scaffolds immobilized with an osteogenic growth peptide for bone tissue regeneration

It is highly desirable for bone tissue engineering scaffolds to have significant osteogenic properties and capability to improve cell growth and thus enhance bone regeneration. In this study, a poly(l-lactic acid) (PLLA) nanofiber scaffold-immobilized osteogenic growth peptide (OGP) was prepared via polydopamine (PDA) coating. X-ray photoelectron spectroscopy (XPS), contact angle measurement, and scanning electron microscopy (SEM) were used to determine the OGP immobilization, hydrophilicity and surface roughness of the samples. The SEM and fluorescence images demonstrate that the PLLA nanofiber scaffolds immobilized with the OGP have excellent cytocompatibility in terms of cell adhesion and proliferation. The ALP activity and the Runx2 and OPN expression results indicated that the PLLA nanofiber scaffolds immobilized with OGP significantly enhanced the osteogenic differentiation and calcium mineralization of hMSCs in vitro. A rat model of critical skull bone defect was selected to evaluate the bone formation capacity of the scaffolds. Micro CT analysis and histological results demonstrated that the PLLA scaffolds immobilized with OGP significantly promoted bone regeneration in critical-sized bone defects. This study verifies that the PLLA scaffold-immobilized OGP has significant potential in bone tissue engineering.

Polydopamine-modified PLLA nanofiber scaffolds immobilized with osteogenic growth peptide were designed and prepared for promoting bone formation.     

Toward improved wound dressings: effects of polydopamine-decorated poly(lactic-co-glycolic acid) electrospinning incorporating basic fibroblast growth factor and ponericin G1

Artificial dressings composed of degradable polymer materials have a wide range of applications in skin repair. The structure and properties, in particular, the antibacterial properties, of the material surface are crucial for biological processes such as cell adhesion, proliferation, and skin regeneration. In this study, we aimed to prepare poly(lactic-co-glycolic acid) (PLGA) nanofiber scaffolds modified by polydopamine using electrospinning technology in order to produce polydopamine-modified degradable PLGA nanocomposites. The polydopamine-PLGA scaffold was endowed with excellent protein adhesion ability through the cross-linking of two biologically active factors, basic fibroblast growth factor (bFGF) and ponericin G1, significantly improving skin repair ability. The electrospun nanofiber scaffold was shown to have a structure similar to that of the natural cell matrix and created a more favorable microenvironment for cell growth. Surface modification by polydopamine dramatically improved the hydrophilicity of the nanofiber scaffold, increasing its ability to absorb active factors and its biocompatibility. The bFGF and ponericin G1 loaded onto the scaffold surface (PDA-PLGA/bFGF/ponericin G1 nanofiber scaffold) strongly promoted the antibacterial and cell proliferation-promoting properties and greatly enhanced the adhesion and proliferation of cells on the scaffold surface. The nanofiber scaffold also promoted wound healing and tissue collagen production in a rat wound healing model. Together, these findings indicate that the polydopamine-PLGA/bFGF/ponericin G1 nanofiber scaffold exhibits good biocompatibility and antibacterial properties, suggesting that it possesses potential value for skin tissue regeneration applications.

Artificial dressings composed of degradable polymer materials have a wide range of applications in skin repair.     

Carbon nanotubes as electrophysiological building blocks for a bioactive cell scaffold through biological assembly to induce osteogenesis

Bio-functional cell scaffolds have great potential in the field of tissue regenerative medicine. In this work, a carbon nanotube (CNT) gel scaffold via specific pairing of functionalized nucleobases was developed for specifically targeted drug delivery and in vitro osteogenesis. The CNT gel scaffold with nano-fibrous architectures was established by Watson–Crick base pairing between thymine and adenine of low molecular weight heparin, respectively. As scaffold precursors, adenine and thymine functionalized heparin derivatives could additionally bind cell growth factors by the affinity interaction. The resulting nano-fibrous gel scaffolds showed excellent mechanical integrity and advanced electro-physiological functions. Potential application of the electrophysiological CNT gel scaffold in bone tissue engineering was confirmed by encapsulation of human adipose-derived stem cells (ASCs). Our results indicate that the electrically conductive networks formed by CNTs within the nano-fibrous framework are the key characteristics of cell scaffolds leading to improved ASC organization and differentiation by an extra electrical stimulus (ES). Specifically, ASCs cultured in bio-electrical gel scaffolds showed ∼4 times higher spontaneous osteogenesis in combination with bone morphogenetic protein 2 (BMP-2), compared to those cultured on pristine hydrogels. This electrophysiological CNT gel scaffold containing BMP-2 exhibited beneficial effects on ASC activity and osteogenetic differentiation, which suggested a promising future for local treatment of bone regeneration.

Bio-functional cell scaffolds have great potential in the field of tissue regenerative medicine.     

Synthesis,characterization and osteogenesis of phosphorylated methacrylamide chitosan hydrogels

Phosphorylated biopolymers can induce mineralization, mimic the process of natural bone formation, and have the potential as scaffolds for bone tissue engineering. 2-Methacryloyloxyethyl phosphorylcholine (MPC), a low cytotoxicity phosphorus source, is mainly applied in vascularization and promoting blood compatibility and has been less researched for bone repair. In this study, phosphorylated methacrylamide chitosan (PMAC) hydrogel was prepared by mixing methacrylamide chitosan (MAC) and different mass of MPC with photoinitiator under UV irradiation. A series characterization tests showed that PMAC hydrogels were successful prepared and had a pretty good mineralization ability. Moreover, human fetal osteoblastic (hFOB) cells cultured on PMAC hydrogels exhibited not only highly viability but also the enhanced ALP activity and calcium deposition. The PMAC hydrogels have great potential in bone tissue engineering applications.

Phosphorylated biopolymers can induce mineralization, mimic the process of natural bone formation, and have the potential as scaffolds for bone tissue engineering.     

Electrospun porous poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/lecithin scaffold for bone tissue engineering

Bone tissue engineering has emerged as a promising restorative strategy for bone reconstruction and bone defect repair. It is challenging to establish an appropriate scaffold with an excellent porous microstructure for bone defects and thereby promote bone repair. In this study, electrospinning as a simple and efficient technology was employed to fabricate a porous poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB) scaffold coated with lecithin. The morphology, phase composition, and physical properties of the electrospun P34HB/lec scaffold were characterized. Meanwhile, cellular behaviors of bone marrow mesenchymal stem cells (BMSCs), including proliferation, adhesion, migration, osteogenic differentiation, and related gene expression, were also investigated. Finally, a rat subcutaneous implant model and a calvarial defect model were used to evaluated the biocompatibility and effect of these scaffolds on bone repair, respectively. The in vitro results demonstrated that these electrospun fibers were interwoven with each other to form the porous P34HB/lec scaffold and the addition of lecithin improved the hydrophilicity of the pure P34HB scaffold, enhanced the efficiency of cell migration, and decreased inflammatory response. Furthermore, the in vivo results showed that P34HB/lec scaffold had excellent biocompatibility, improved the vascularization, and promoted the bone regeneration. All these results indicated that nanofibers of P34HB scaffolds in combination with the lecithin could exert a synergistic effect on promoting osteogenesis and regeneration of bone defects; thus, the P34HB scaffold with lecithin showed great application potential for bone tissue engineering.

An electrospun P34HB scaffold was prepared and coated with lecithin. As a scaffold for bone tissue engineering, the P34HB/lec scaffold could promote proliferation and osteogenesis of BMSCs in vitro, and also accelerate bone regeneration in vivo.     

Study on antibacterial properties and cytocompatibility of EPL coated 3D printed PCL/HA composite scaffolds

Biomaterial scaffolds play a critical role in bone tissue engineering. Moreover, 3D printing technology has enormous advantage in the manufacture of bioengineering scaffolds for patient-specific bone defect treatments. In order to provide an aseptic environment for bone regeneration, ε-poly-l-lysine (EPL), an antimicrobic cationic polypeptide, was used for surface modification of 3D printed polycaprolactone/hydroxyapatite (PCL/HA) scaffolds which were fabricated by fused deposition modeling (FDM) technology. The scaffold morphology and micro-structure were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and transform infrared spectroscopy (FT-IR). The release profile surface roughness, open porosity, and mechanical properties of the scaffolds were evaluated. Cell adhesion, proliferation, differentiation potential and antibacterial properties were also examined. As a result, 3D printed PCL/HA scaffolds with interconnected pores showed a slightly rough surface and improved mechanical properties due to adding hydroxyapatite (HA) particles. After being modified by EPL, favorable biocompatibility and osteoconductivity of ε-poly-l-lysine/polycaprolactone/hydroxyapatite (EPL/PCL/HA) scaffolds were observed. Moreover, antibacterial activity of the EPL/PCL/HA scaffolds was apparent. As a consequence, the EPL/PCL/HA scaffolds had great potential for bone regeneration and prevention of infections. This would yield a patient-specific bioactive and antibacterial composite scaffold for advanced bone tissue engineering applications.

Biomaterial scaffolds play a critical role in bone tissue engineering.     

Biomaterials for bone tissue engineering scaffolds: a review

Bone tissue engineering has been continuously developing since the concept of “tissue engineering” has been proposed. Biomaterials that are used as the basic material for the fabrication of scaffolds play a vital role in bone tissue engineering. This paper first introduces a strategy for literature search. Then, it describes the structure, mechanical properties and materials of natural bone and the strategies of bone tissue engineering. Particularly, it focuses on the current knowledge about biomaterials used in the fabrication of bone tissue engineering scaffolds, which includes the history, types, properties and applications of biomaterials. The effects of additives such as signaling molecules, stem cells, and functional materials on the performance of the scaffolds are also discussed.

Bone tissue engineering has been continuously developing since the concept of “tissue engineering” has been proposed. Biomaterials, as the basic material for the fabrication of scaffolds, play a vital role in bone tissue engineering.     

Bioceramic akermanite enhanced vascularization and osteogenic differentiation of human induced pluripotent stem cells in 3D scaffolds in vitro and vivo

A growing number of studies suggest that the modulation of cell differentiation by biomaterials is critical for tissue engineering. In previous work, we demonstrated that human induced pluripotent stem cells (iPSCs) are remarkably promising seed cells for bone tissue engineering. In addition, we found that the ionic products of akermanite (Aker) are potential inducers of osteogenic differentiation of iPSCs. Furthermore, composite scaffolds containing polymer and bioceramics have more interesting properties compared to pure bioceramic scaffolds for bone tissue engineering. The characteristic of model biomaterials in bone tissue engineering is their ability to control the osteogenic differentiation of stem cells and simultaneously induce the angiogenesis of endothelia cells. Thus, this study aimed at investigating the effects of poly(lactic-co-glycolic acid)/Aker (PLGA-Aker) composite scaffolds on angiogenic and osteogenic differentiation of human iPSCs in order to optimize the scaffold compositions. The results from Alizarin Red S staining, qRT-PCR analysis of osteogenic genes (BMP2, RUNX2, ALP, COL1 and OCN) and angiogenic genes (VEGF and CD31) demonstrated that PLGA/Aker composite scaffolds containing 10% Aker exhibited the highest stimulatory effects on the osteogenic and angiogenic differentiation of human iPSCs among all scaffolds. After the scaffolds were implanted in nu/nu mice subcutaneous pockets and calvarial defects, H&E staining, BSP immunostaining, qRT-PCR analysis and micro-CT analysis (BMD, BV/TV) indicated that PLGA + 10% Aker scaffolds enhanced the vascularization and osteogenic differentiation of human iPSCs and stimulated the repair of bone defects. Taken together, our work indicated that combining scaffolds containing silicate bioceramic Aker and human iPSCs is a promising approach for the enhancement of bone regeneration.

Bioceramics akermanite enhanced vascularization and osteogenic differentiation of human iPSCs in 3D scaffolds in vitro and vivo.     

Sodium alginate/collagen composite multiscale porous scaffolds containing poly(ε-caprolactone) microspheres fabricated based on additive manufacturing technology

Biocompatible porous scaffolds with adjustable pore structures, appropriate mechanical properties and drug loading properties are important components of bone tissue engineering. In this work, biocompatible sodium alginate (SA)/collagen (Col) multiscale porous scaffolds containing poly(ε-caprolactone) microspheres (Ms-PCL) have been facilely fabricated based on 3D extrusion printing of the pre-crosslinked composite hydrogels. The prepared composite hydrogels can be 3D extrusion printed into porous scaffolds with different designed shapes and adjustable pore structures. The hydroxyapatite (HAP) nanoparticles have been added into the SA/Col hydrogels to achieve stress dispersion and form double crosslinking networks. SA-Ca²⁺ crosslinking networks and Col–genipin (GP) crosslinking networks have been constructed to improve the mechanical properties of the scaffolds (about 2557 kPa of compressive stress at 70% strain), and reduce the swelling rate and degradation rate of SA/Col scaffolds. Moreover, the SA/Col hydrogels contain hydrophobic antibacterial drug enrofloxacin loaded Ms-PCL, and in vitro drug release research shows a sustained-release function of porous scaffolds, indicating the potential application of SA/Col porous scaffolds as drug carriers. In addition, the antibacterial experiments show that the composite scaffolds display a distinguished and long-term antibacterial activity against Escherichia coli and Staphylococcus aureus. Furthermore, mouse bone mesenchymal stem cells (mBMSCs) are seeded on the SA/Col composite scaffolds, and an in vitro biocompatibility experiment shows that the mBMSCs can adhere well on the composite scaffolds, which indicate that the fabricated composite scaffolds are biocompatible. In short, all of the above results suggest that the biocompatible SA/Col composite porous scaffolds have enormous application and potential in bone tissue engineering.

Biocompatible porous scaffolds with adjustable pore structures, appropriate mechanical properties and drug loading properties are important components of bone tissue engineering.     

A three-dimensional hydroxyapatite/polyacrylonitrile composite scaffold designed for bone tissue engineering

In recent years, various composite scaffolds based on hydroxyapatite have been developed for bone tissue engineering. However, the poor cell survival micro-environment is still the major problem limiting their practical applications in bone repairing and regeneration. In this study, we fabricated a class of fluffy and porous three-dimensional composite fibrous scaffolds consisting of hydroxyapatite and polyacrylonitrile by employing an improved electrospinning technique combined with a bio-mineralization process. The fluffy structure of the hydroxyapatite/polyacrylonitrile composite scaffold ensured the cells would enter the interior of the scaffold and achieve a three-dimensional cell culture. Bone marrow mesenchymal stem cells were seeded into the scaffolds and cultured for 21 days in vitro to evaluate the response of cellular morphology and biochemical activities. The results indicated that the bone marrow mesenchymal stem cells showed higher degrees of growth, osteogenic differentiation and mineralization than those cultured on the two-dimensional hydroxyapatite/polyacrylonitrile composite membranes. The obtained results strongly supported the fact that the novel three-dimensional fluffy hydroxyapatite/polyacrylonitrile composite scaffold had potential application in the field of bone tissue engineering.

A fluffy and porous (3D) HA composite fibrous scaffold was fabricated by employing an improved electrospinning technique combined with a bio-mineralization process.     

Enhanced regeneration of bone defects using sintered porous Ti6Al4V scaffolds incorporated with mesenchymal stem cells and platelet-rich plasma

A new highly controlled powder sintering technique was used for the fabrication of a porous Ti6Al4V scaffold. The platelet-rich plasma (PRP) was prepared using whole blood. The PRP was used as a cell carrier to inject bone marrow mesenchymal stem cells (MSC) into the pores of the Ti6Al4V scaffold in the presence of calcium chloride and thrombin, and then the composite construct of porous Ti6Al4V loaded with PRP gel and MSC was obtained. The bare Ti6Al4V scaffold and the Ti6Al4V scaffold loaded with MSC were used as controls. The characteristics and mechanical properties of the scaffold, and the biological properties of the constructs were evaluated by a series of in vitro and in vivo experiments. The results show that the sintered porous Ti6Al4V has good biocompatibility, and high porosity and large pore size, which can provide sufficient space and sufficient mechanical support for the growth of cells and bones without an obvious stress shielding effect. However, Ti6Al4V/MSC/PRP showed a significantly higher cell proliferation rate, faster bone growth speed, more bone ingrowth, and higher interfacial strength. Therefore, the porous Ti6Al4V scaffolds incorporated with MSC and PRP may be more effective at enhancing bone regeneration, and is expected to be used for bone defect repair.

Porous Ti6AI4V scaffolds incorporated with MSC and PRP are more effective in enhancing the bone regeneration.     

The enhanced angiogenesis effect of VEGF-silk fibroin nanospheres-BAMG scaffold composited with adipose derived stem cells in a rabbit model

We report a study to determine whether a vascular endothelial growth factor (VEGF)-silk fibroin (SF) nanospheres-bladder acellular matrix graft (BAMG) scaffold composited with adipose derived stem cells (ADSCs) could enhance angiogenesis in bladder regeneration in rabbits. Rabbit ADSCs were isolated and identified by flow cytometry. The morphology and release behaviour of VEGF-SF nanospheres were detected. After the composite scaffolds were successfully used in bladder reconstruction, the bladder capacity, H&E staining and immunohistochemical staining were studied at different time points. ADSCs exerts high expression rates of CD29, CD90, and CD44, accompanied with low expression rates of CD34 and CD45. SF nanospheres with diameters of 200–1000 nm were prepared to load VEGF, and they contributed to maintain the release of VEGF. The reconstructed bladder with VEGF-SF nanospheres-BAMG plus ADSCs had more regular smooth muscle tissue and blood vessels. Moreover, instead of differentiating into epithelial or vascular endothelial cells, ADSCs may be more likely to provide additional cytokines to enhance angiogenesis in the bladder regeneration process. The tissue engineered bladder constructed by BAMG modified by VEGF-SF nanospheres possessed high bio-compatibility and an enhanced angiogenesis effect, and could be used as an ideal biological material to repair bladder defects after being composited with ADSCs.

The adipose derived stem cells (ADSCs) was composited with vascular endothelial growth factor (VEGF)-silk fibroin (SF) nanospheres-bladder acellular matrix graft (BAMG) scaffold to repair bladder defect in rabbits.     

Anisotropic architecture and electrical stimulation enhance neuron cell behaviour on a tough graphene embedded PVA: alginate fibrous scaffold

Tough scaffolds comprised of aligned and conductive fibers are promising for peripheral nerve regeneration due to their unique mechanical and electrical properties. Several studies have confirmed that electrical stimulation can control the axonal extension in vitro. However, the stimulatory effects of scaffold architecture and electrical stimulation have not yet been investigated in detail. Here, we assessed a comparison between aligned and random fibers made of graphene (Gr) embedded sodium alginate (SA) polyvinyl alcohol (PVA) (Gr-AP scaffolds) for peripheral nerve engineering. The effects of applied electrical stimulation and orientation of the fabricated fibers on the in vitro attachment, alignment, and proliferation of PC12 cells (a rat neuronal cell line) were investigated. The results revealed that the aligned fibrous Gr-AP scaffolds closely mimicked the anisotropic structure of the native sciatic nerve. Aligned fibrous Gr-AP scaffolds significantly improved mechanical properties as well as cell-scaffold integration compared to random fibrous scaffolds. In addition, electrical stimulation significantly improved PC12 cell proliferation. In summary, our findings revealed that aligned fibrous Gr-AP scaffolds offered superior mechanical characteristics and structural properties that enhanced neural cell–substrate interactions, resulting in a promising construct for nerve tissue regeneration.

Tough scaffolds comprised of aligned and conductive fibers are promising for peripheral nerve regeneration due to their unique mechanical and electrical properties.     

3D printable SiO2 nanoparticle ink for patient specific bone regeneration

Sodium alginate and gelatin are biocompatible & biodegradable natural polymer hydrogels, which are widely investigated for application in tissue engineering using 3D printing and 3D bioprinting fabrication techniques. The major challenge of using hydrogels for tissue fabrication is their lack of regeneration ability, uncontrolled swelling, degradation and inability to hold 3D structure on their own. Free hydroxyl groups on the surface of SiO₂ nanoparticles have the ability to chemically interact with alginate–gelatin polymer network, which can be explored to achieve the above parameters. Hence validating the incorporation of SiO₂ nanoparticles in a 3D printable hydrogel polymer network, according to the patient''s critical defects has immense scope in bone tissue engineering. In this study, SiO₂ nanoparticles are loaded into alginate–gelatin composite hydrogels and chemically crosslinked with CaCl₂ solution. The effect of SiO₂ nanoparticles on the viscosity, swelling, degradation, compressive modulus (MPa), biocompatibility and osteogenic ability were evaluated on lyophilized scaffolds and found to be desirable for bone tissue engineering. A complex irregular patient-specific virtual defect was created and the 3D printing process to fabricate such structures was evaluated. The 3D printing of SiO₂ nanoparticle hydrogel composite ink to fabricate a bone graft using a patient-specific virtual defect was successfully validated. Hence this type of hydrogel composite ink has huge potential and scope for its application in tissue engineering and nanomedicine.

3D printing of a complex and irregular virtual defect using SiO₂ nanoparticle and hydrogel composite ink for patient specific defect fabrication.