Electrospinning is a simple and efficient process for producing sub-micron fibres. However, the process has many variables, and their effects on the non-woven mesh of fibres is complex. In particular, the effects on the mechanical properties of the fibre meshes are poorly understood. This paper conducts a parametric study, where the concentration and bloom strength of the gelatin solutions are varied, while all electrospinning process parameters are held constant. The effects on the fibrous meshes are monitored using scanning electron microscopy and mechanical testing under uniaxial tension. Mesh mechanical properties are relatively consistent, despite changes to the solutions, demonstrating the robustness of electrospinning. The gel strength of the solution is shown to have a statistically significant effect on the morphology, stiffness and strength of the meshes, while the fibre diameter has surprisingly little influence on the stiffness of the meshes. This experimental finding is supported by finite element analysis, demonstrating that the stiffness of the meshes is controlled by the volume fraction, rather than fibre diameter. Our results demonstrate the importance of understanding how electrospinning parameters influence the pore size of the meshes, as controlling fibre diameter alone is insufficient for consistent mechanical properties.
Sodium alginate/hydroxyapatite/Nano cellulose (SA/HA/NC) nanocomposite films that possess good biocompatibility for bone tissue engineering are prepared by a simple solution casting. HA is one of the most frequently used bioceramic materials to achieve a high biocompatibility. The bionanocomposite films are analysed by XRD, SEM, EDAX and FTIR studies. XRD confirms the existence of fillers in the polymer. FTIR spectrum shows the different functional modes in the bionanocomposite films. The morphology of fillers and bionanocomposite films are obtained through SEM. The inclusion of NC with different concentrations into the biopolymer film improves the tensile strength. As a result, the loading of 5 wt % of NC and 10 wt% of HA in the SA polymer shows high tensile strength when compared to the pure SA, SA filled with 10 wt% of HA and SA loaded with 10 wt% of HA and inclusion of NC (0.5 and 2.5 wt%). The tensile strength (TS) of bionanocomposite film with 10 wt % of HA is increased by 17%. TS of bionanocomposite film with 0.5 and 2.5 wt% of NC is increased by 177 and 277%, whereas TS of bionanocomposite film loaded 5 wt% of NC is increased by 331%. The swelling, biodegradation and biomineralization tests suggest that this bionanocomposite films are hopeful biomaterials for bone tissue engineering.
The full-thickness skin wound is a common skin complication affecting millions of people worldwide. Delayed treatment of this condition causes the loss of skin function and integrity that could lead to the development of chronic wounds or even death. This study was aimed to develop a rapid wound treatment modality using ovine tendon collagen type I (OTC-I) bio-scaffold with or without noncultured skin cells. Genipin (GNP) and carbodiimide (EDC) were used to cross-link OTC-I scaffold to improve the mechanical strength of the bio-scaffold. The physicochemical, biomechanical, biodegradation, biocompatibility, and immunogenicity properties of OTC-I scaffolds were investigated. The efficacy of this treatment approach was evaluated in an in vivo skin wound model. The results demonstrated that GNP cross-linked OTC-I scaffold (OTC-I_GNP) had better physicochemical and mechanical properties compared with EDC cross-linked OTC-I scaffold (OTC-I_EDC) and noncross-link OTC-I scaffold (OTC-I_NC). OTC-I_GNP and OTC-I_NC demonstrated no toxic effect on cells as it promoted higher cell attachment and proliferation of both primary human epidermal keratinocytes and human dermal fibroblasts compared with OTC-I_EDC. Both OTC-I_GNP and OTC-I_NC exhibited spontaneous formation of bilayer structure in vitro. Immunogenic evaluation of OTC-I scaffolds, in vitro and in vivo, revealed no sign of immune response. Finally, implantation of OTC-I_NC and OTC-I_GNP scaffolds with noncultured skin cells demonstrated enhanced healing with superior skin maturity and microstructure features, resembling native skin in contrast to other treatment (without noncultured skin cells) and control group. The findings of this study, therefore, suggested that both OTC-I scaffolds with noncultured skin cells could be promising for the rapid treatment of full-thickness skin wound.
Previous studies suggested telomerase activity as a determinant of cell replicative capacity by delaying cell senescence. This study aimed to evaluate the feasibility of adopting telomerase activity as a selection criterion for in vitro expanded skin cells before autologous transplantation. Fibroblasts and keratinoctyes were derived from the same consenting patients aged 9-69 years, and cultured separately in serum-supplemented and serum-free media, respectively. Telomerase activity of fresh and cultured cells were measured and correlated with cell growth rate, donor age and passage number. The results showed that telomerase activity and cell growth were independent of donor age for both cell types. Telomerase was expressed in freshly digested epidermis and dermis and continued expressing in vitro. Keratinocytes consistently showed 3-12 folds greater telomerase activity than fibroblast both in vivo and in vitro. Conversely, growth rate for fibroblast exceeded that of keratinocyte. Telomerase activity decreased markedly at Passage 6 for keratinocytes and ceased by Passage 3 for fibroblasts. The decrease or cessation of telomerase activity coincided with senescence for keratinocyte but not for fibroblast, implying a telomerase-regulated cell senescence for the former and hence a predictor of replicative capacity for this cell type. Relative telomerase activity for fibroblasts from the younger age group was significantly higher than that from the older age group; 69.7% higher for fresh isolates and 31.1% higher at P0 (p<0.05). No detectable telomerase activity was to be found at later subcultures for both age groups. Similarly for keratinocytes, telomerase activity in the younger age group was significantly higher (p<0.05) compared to that in the older age group; 507.7% at P0, 36.8% at P3 and the difference was no longer significant at P6. In conclusion, the study provided evidence that telomerase sustained the proliferation of keratinocytes but not fibroblasts. Telomerase activity is an important criterion for continued survival and replication of keratinocytes, hence its positive detection before transplantation is desirable. Inferring from our results, the use of keratinocytes from Passage 3 or lesser for construction of skin substitute or cell-based therapy is recommended owing to their sustained telomerase expression.
Polyhydroxyalkanoates (PHAs) have attracted the attention of academia and industry because of their plastic-like properties and biodegradability. However, practical applications as a commodity material have not materialized because of their high production cost and unsatisfactory mechanical properties. PHAs are also believed to have high-value applications as an absorbable biomaterial for tissue engineering and drug-delivery devices because of their biocompatibility. However, research in these areas is still in its very early stages. The main problem faced by proponents of PHAs is the lack of a niche area where PHAs will be the most desired material in terms of its function during use rather than because of its eco-friendly virtues after use. Here, we report on the oil-absorbing property of PHA films and its potential applications. By comparing with some of the existing commercial products, the potential application of PHAs as cosmetic oil-blotting films is revealed for the first time. Besides having the ability to rapidly absorb and retain oil, PHA films also have a natural oil-indicator property, showing obvious changes in opacity following oil absorption. Surface analysis revealed that the surface structures such as porosity and smoothness exert great influence on the rapid oil-absorption properties of the PHA films. These newly discovered properties could be exploited to create a niche area for the practical applications of PHAs.
In vitro production of sperm is a desirable idea for fertility preservation in azoospermic men and prepubertal boys suffering from cancer. In this study, a biocompatible porous scaffold based on a triad mixture of silk fibroin (SF), alginate (Alg), and laminin (LM) is developed to facilitate the differentiation of mouse spermatogonia stem cells (SSCs). Following SF extraction, the content is analyzed by SDS-PAGE and stable porous 3D scaffolds are successfully prepared by merely Alg, SF, and a combination of Alg-SF, or Alg-SF-LM through freeze-drying. Then, the biomimetic scaffolds are characterized regarding the structural and biological properties, water absorption capacity, biocompatibility, biodegradability, and mechanical behavior. Neonatal mice testicular cells are seeded on three-dimensional scaffolds and their differentiation efficiency is evaluated using real-time PCR, flow cytometry, immunohistochemistry. Blend matrices showed uniform porous microstructures with interconnected networks, which maintained long-term stability and mechanical properties better than homogenous structures. Molecular analysis of the cells after 21 days of culture showed that the expression of differentiation-related proteins in cells that are developed in composite scaffolds is significantly higher than in other groups. The application of a composite system can lead to the differentiation of SSCs, paving the way for a novel infertility treatment landscape in the future.
Tissue regeneration is a rapidly growing field providing a beacon of hope in the field of restorative and endodontics. Root canal treatment involves the removal of pulp tissue and replacement by an inorganic materials where as regenerative endodontics deals with replacement with healthy pulp to revitalize the teeth .Research in the field of tissue engineering and material science have lead to significant progress but still is plague with lots of drawbacks and failures, hence it is still not being adapted as routine clinical procedures .The purpose of this article is to review the advances made in regenerative endodontics and the future scopes.
Introduction: Oral cancer is the sixth most common malignancy in the world. It is a major concern in Southeast Asia primarily due to betel quid chewing, smoking, and alcohol consumption. In Malaysia, oral cancer related cases accounts for 1.55% of the cause of deaths. Despite recent advances in cancer diagnoses and therapies, the survival rate of oral cancer patients only reached 50% in the last few decades. Tissue engineering (TE) principles may pro-vide new technology platforms to study mechanisms of angiogenesis and tumour cell growth as well as potentially tumour cell spreading in cancer research. The use of biomaterial, appropriate cell source and proper signalling mol-ecules are vital components of TE. Collagen biomaterial are widely used scaffold or membrane in oral application. Nevertheless, no review has been performed on the its usage for the study of oral cancer. This study aimed to sys-tematically review the use of collagen scaffold in oral cancer application. Methods: Research articles were searched using Scopus, Pubmed and Web of Science (WOS) databases. The keywords were limited to “collagen membrane OR collagen scaffold” AND “oral cancer”. Results: Initial search yielded 61 papers (Scopus:37, Pubmed: 12, WOS: 12). Further scrutinization of the papers based on the inclusion criteria resulted total of 3 papers. Two of the papers used collagen membrane for regeneration of oral mucosal defect and increment of alveolar ridge height post-surgery. The remaining paper utilize collagen biomaterial as scaffold for the culture of adenoid cystic carcinoma (ACC) cells. All papers reported significant role of collagen biomaterial in terms of tissue formation, healing scaffold and cellular proliferation. Conclusion: Collagen utilization as biomaterial offers potential use for regeneration of oral related structures as well providing useful model for therapeutics anti-cancer research.
Introduction: Tissue-engineered oral mucosa (TEOM) is increasingly being used to model oral mucosal diseases and to assess drug toxicity. Current TEOM models are constructed using normal oral fibroblasts (NOF) contained within a hydrogel matrix with normal oral keratinocytes (NOK) cultured on top. NOK are not commercially available and suffer from donor-to-donor variability. Therefore, oral mucosal models based on immortalised keratinocytes may offer advantages over NOK-based models. The objective of this study was to construct and characterise the TEOM developed using TERT2-immortalised oral keratinocyte (FNB6) cells and validate its similarity to normal oral muco-sal tissue. Methods: TEOM were constructed by culturing FNB6 cells on top of a NOF-populated collagen type-1 hydrogel in tissue culture transwell inserts cultured at an air-to-liquid interface and collected at 14 day. TEOM were subjected to morphological (H&E and PAS), ultrastructural (TEM) and immunohistological (Ki-67, cytokeratin 14 and E-cadherin) analysis. Results: Histologically TEOM mimicked native oral mucosa displaying a stratified epithelium, fibroblast-containing connective tissue and basement membrane. Furthermore, TEM confirmed the presence of des-mosomes and hemi-desmosomes in the epithelium. IHC revealed expression of differentiation markers (cytokeratin 14), proliferation (Ki-67), cell adhesion (E-cadherin). Conclusion: FNB6 mucosal models able to mimic native oral mucosa structure. It has potential for drug delivery and toxicity evaluation, and replacing models based on NOK where access to primary cells is limited.
Presently, there is no specific federal legislation governing articular cartilage tissue engineering (ACTE) experimenta- tion practices in Malaysia. However, there are related regulations and guidelines provided by government agencies to oversee and guide such practices. The rules and regulations provided in the documents have the essential aim of safeguarding public health through ensuring that non-clinical studies reach a certain quality, efficient and safe for hu- man use. There are themes identified when scrutinising relevant documents which includes, the need for authorised personnel and the establishment of facilities in conducting such experiments, the aspect of cell-scaffold construct development, the use of human materials, the aspect of biosafety, animal care and use during the experiments, and considerations on the impact on the environment. The individual laboratory or facility shall adopt and adapt these standards as deemed appropriate by the ACTE researchers to ensure that non-clinical studies are conducted in a proper and ethical manner.
Angiogenicity is one of the essential components to enable tissue function. It is important to develop a construct that would help in catering oxygen and nutrient to the engineered tissue area. Thus, this study aims to investigate the attachment, spreading and growth of stem cells from human exfoliated deciduous teeth (SHED) on human AM (HAM) with or without vascular endothelial growth factor (VEGF) using scanning electron microscope (SEM), and indirectly see the potential of the HAM as a scaffold to promote angiogenic micro-environment. Since day 1, there were continuous changes of the cell morphology until day 28, SHED treated with VEGF seemed to change its shape from fibroblast-like into a round-shape cell, similar structure as an endotheliallike cell. The structures of filopodia-like were also observed on the treated SHED. SHED without VEGF treatment showed only normal morphological growth on HAM. VEGF is a protein produced to stimulate angiogenesis, and is believed to contribute to the morphological changes of SHED seeded on HAM. This indicates that HAM could be used as a scaffold to allow SHED differentiation into endothelial-like cells with the induction of VEGF.
Porous ceramic scaffolds are widely studied in the tissue engineering field due to their potential in medical applications as bone substitutes or as bone-filling materials. In this study, porous hydroxyapatite (HA) was produced via polymer replication method. Polyurethane (PU) sponge was selected as the template and synthetic binder, polyvinyl alcohol (PVA) was used in this study. Fixed formulation of HA powder, distilled water and PVA (40:60:3) were prepared and stirred at a constant 4 hours time. PU sponges with 30 ppi and 60 ppi size were cut and impregnated in slurry using vacuum and roller infiltration methods. The microstructures were observed by using field emission scanning electron microscope (FESEM). The results obtained indicate that vacuum infiltration method and 60 ppi template pore size exhibited the highest compressive strength with moderate average strut thickness and lowest average pore size compared to samples produced by roller infiltration method at different template pore size.
Meniscectomy is the most common surgery in orthopaedics. The absence of meniscal tissue might be related to irreversible damage to the articular cartilage. Meniscal replacement is a tissue-engineering technique for post-meniscectomy syndrome. Its success depends on the implant integration which was vastly proven in animal model studies. Histological evidence is hard to obtain in humans due to ethical issues. We report a clinical case in which a collagen scaffold meniscal implant was harvested six months after implantation due to mechanical failure. Histological analysis was performed revealing vascularisation not only of the peripheral attachment of the implant but also on the anterior horn. These morphologic findings demonstrate that this implant allows the colonisation by precursor cells and vessels, leading to the formation of a fully functional tissue. This present report is one of the few independent reports of scaffold biological integration in the literature.
Purpose:To investigate the feasibilty of using processed human amniotic membrane (HAM) to support the attachment and proliferation of chondrocytes in vitro which it turn can be utilised as a cell delivery vehicle in tissue engineering applications. Methods: Fresh HAM obtained from patients undergoing routine elective ceasarean sections was harvested., processed and dried using either freez drying (FD) or air drying (AD) methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrolytes were seeded on processsed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microcospy. Results: Processed HAM appeared to allow cell attachment when implanted with chrondocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. Conclusion: Prelimanary results show that processed HAM chondrocyte attachment and proliferation.
Hydroxyapatite (HA) has been widely used as a scaffold in tissue engineering. HA possesses high mechanical stress and exhibits particularly excellent biocompatibility owing to its similarity to natural bone. Nonetheless, this ceramic scaffold has limited applications due to its apparent brittleness. Therefore, this had presented some difficulties when shaping implants out of HA and for sustaining a high mechanical load. Fortunately, these drawbacks can be improved by combining HA with other biomaterials. Starch was heavily considered for biomedical device applications in favor of its low cost, wide availability, and biocompatibility properties that complement HA. This review provides an insight into starch/HA composites used in the fabrication of bone tissue scaffolds and numerous factors that influence the scaffold properties. Moreover, an alternative characterization of scaffolds via dielectric and free space measurement as a potential contactless and nondestructive measurement method is also highlighted.
The four heart valves represented in the mammalian hearts are responsible for maintaining unidirectional, non-hinder blood flow. The heart valve leaflets synchronically open and close approximately 4 million times a year and more than 3 billion times during the life. Valvular heart dysfunction is a significant cause of morbidity and mortality around the world. When one of the valves malfunctions, the medical choice is may be to replace the original valves with an artificial one. Currently, the mechanical and biological artificial valves are clinically used with some drawbacks. Tissue engineering heart valve concept represents a new technique to enhance the current model. In tissue engineering method, a three-dimensional scaffold is fabricated as the template for neo-tissue development. Appropriate cells are seeded to the matrix in vitro. Various approaches have been investigated either in scaffold biomaterials and fabrication techniques or cell source and cultivation methods. The available results of ongoing experiments indicate a promising future in this area (particularly in combination of bone marrow stem cells with synthetic scaffold), which can eliminate the need for lifelong anti-coagulation medication, durability and reoperation problems.
Magnetic nanofibers are composed of good dispersion of magnetic nanoparticles along an organic material. Magnetic nanofibers are potentially useful for composite reinforcement, bio-medical and tissue engineering. Nanofibers with the thinner diameter have to result in higher rigidity and tensile strength due to better alignments of lamellae along the fiber axis. In this study, the performance of electrospinning process was explained using response surface methodology (RSM) during fabrication of magnetic nanofibers using polyvinyl alcohol (PVA) as a shelter for (γ-Fe2O3) nanoparticles where the parameters investigated were flow rate, applied voltage, distance between needle and collector and collector rotating speed. The response variable was diameter distribution. The two parameters flow rate and applied voltage in primary evaluation were distinguished as significant factors. Central composite design was applied to optimize the variable of diameter distribution. Quadratic estimated model developed for diameter distribution indicated the optimum conditions to be flow rate of 0.25 ml/h at voltage of 45 kV while the distance and rotating speed are at 8 cm and 1500 rps respectively. The obtained model was verified successfully by the confirmation experiments.
The actual in vivo tissue scaffold offers a three-dimensional (3D) structural support along with a nano-textured surfaces consist of a fibrous network in order to deliver cell adhesion and signaling. A scaffold is required, until the tissue is entirely regenerated or restored, to act as a temporary ingrowth template for cell proliferation and extracellular matrix (ECM) deposition. This review depicts some of the most significant three dimensional structure materials used as scaffolds in various tissue engineering application fields currently being employed to mimic in vivo features. Accordingly, some of the researchers' attempts have envisioned utilizing graphene for the fabrication of porous and flexible 3D scaffolds. The main focus of this paper is to evaluate the topographical and topological optimization of scaffolds for tissue engineering applications in order to improve scaffolds' mechanical performances.
Biodegradable elastomers have clinical applicability due to their biocompatibility, tunable degradation and elasticity. The addition of bioactive glasses to these elastomers can impart mechanical properties sufficient for hard tissue replacement. Hence, a composite with a biodegradable polymer matrix and a bioglass filler can offer a method of augmenting existing tissue. This article reviews the applications of such composites for skeletal augmentation.
Recently, porous magnesium and its alloys are receiving great consideration as biocompatible and biodegradable scaffolds for bone tissue engineering application. However, they presented poor antibacterial performance and corrosion resistance which limited their clinical applications. In this study, Mg-Zn (MZ) scaffold containing different concentrations of tetracycline (MZ-xTC, x = 1, 5 and 10%) were fabricated by space holder technique to meet the desirable antibacterial activity and corrosion resistance properties. The MZ-TC contains total porosity of 63-65% with pore sizes in the range of 600-800 μm in order to accommodate bone cells. The MZ scaffold presented higher compressive strength and corrosion resistance compared to pure Mg scaffold. However, tetracycline incorporation has less significant effect on the mechanical and corrosion properties of the scaffolds. Moreover, MZ-xTC scaffolds drug release profiles show an initial immediate release which is followed by more stable release patterns. The bioactivity test reveals that the MZ-xTC scaffolds are capable of developing the formation of HA layers in simulated body fluid (SBF). Next, Staphylococcus aureus and Escherichia coli bacteria were utilized to assess the antimicrobial activity of the MZ-xTC scaffolds. The findings indicate that those scaffolds that incorporate a high level concentration of tetracycline are tougher against bacterial organization than MZ scaffolds. However, the MTT assay demonstrates that the MZ scaffolds containing 1 to 5% tetracycline are more effective to sustain cell viability, whereas MZ-10TC shows some toxicity. The alkaline phosphatase (ALP) activity of the MZ-(1-5)TC was considerably higher than that of MZ-10TC on the 3 and 7 days, implying higher osteoblastic differentiation. All the findings suggest that the MZ-xTC scaffolds containing 1 to 5% tetracycline is a promising candidate for bone tissue healing due to excellent antibacterial activity and biocompatibility.