Bioinks and bioprinting technologies to make heterogeneous and biomimetic tissue constructs.

The native tissues are complex structures consisting of different cell types, extracellular matrix materials, and biomolecules. Traditional tissue engineering strategies have not been able to fully reproduce biomimetic and heterogeneous tissue constructs because of the lack of appropriate biomaterials and technologies. However, recently developed three-dimensional bioprinting techniques can be leveraged to produce biomimetic and complex tissue structures. To achieve this, multicomponent bioinks composed of multiple biomaterials (natural, synthetic, or hybrid natural-synthetic biomaterials), different types of cells, and soluble factors have been developed. In addition, advanced bioprinting technologies have enabled us to print multimaterial bioinks with spatial and microscale resolution in a rapid and continuous manner, aiming to reproduce the complex architecture of the native tissues. This review highlights important advances in heterogeneous bioinks and bioprinting technologies to fabricate biomimetic tissue constructs. Opportunities and challenges to further accelerate this research area are also described.

Three-dimensional printing of metals for biomedical applications.

Three-dimensional (3D) printing technology has received great attention in the past decades in both academia and industry because of its advantages such as customized fabrication, low manufacturing cost, unprecedented capability for complex geometry, and short fabrication period. 3D printing of metals with controllable structures represents a state-of-the-art technology that enables the development of metallic implants for biomedical applications. This review discusses currently existing 3D printing techniques and their applications in developing metallic medical implants and devices. Perspective about the current challenges and future directions for development of this technology is also presented.

Improvement and characterization of the adhesion of electrospun PLDLA nanofibers on PLDLA-based 3D object substrates for orthopedic application.

Intensive research has demonstrated the clear biological potential of electrospun nanofibers for tissue regeneration and repair. However, nanofibers alone have limited mechanical properties. In this study we took poly(L-lactide-co-D-lactide) (PLDLA)-based 3D objects, one existing medical device (interference screws) and one medical device model (discs) as examples to form composites through coating their surface with electrospun PLDLA nanofibers. We specifically investigated the effects of electrospinning parameters on the improvement of adhesion of the electrospun nanofibers to the PLDLA-based substrates. To reveal the adhesion mechanisms, a novel peel test protocol was developed for the characterization of the adhesion and delamination phenomenon of the nanofibers deposited to substrates. The effect of incubation of the composites under physiological conditions on the adhesion of the nanofibers has also been studied. It was revealed that reduction of the working distance to 10 cm resulted in deposition of residual solvent during electrospinning of nanofibers onto the substrate, causing fiber-fiber bonding. Delamination of this coating occurred between the whole nanofiber layer and substrate, at low stress. Fibers deposited at 15 cm working distance were of smaller diameter and no residual solvent was observed during deposition. Delamination occurred between nanofiber layers, which peeled off under greater stress. This study represents a novel method for the alteration of nanofiber adhesion to substrates, and quantification of the change in the adhesion state, which has potential applications to develop better medical devices for orthopedic tissue repair and regeneration.

Chondrogenic potential of electrospun nanofibres for cartilage tissue engineering.

Articular cartilage has a heterogeneous structure, comprising elongated cells at the articulating surface and rounded cells elsewhere. This feature poses a complex challenge when fabricating 3D tissue engineering scaffolds able to mimic the native extracellular matrix (ECM) of cartilage for tissue repair and regeneration. Nanofibre scaffolds can provide an ECM-like structure, but are mechanically weak and typically have subcellular pore geometries. In this study, the use of poly(L,D-lactide) (PLDLA) nanofibre coatings on PLDLA microfibres or films (nanofibre composites) to influence bovine chondrocyte behaviour was investigated. It was demonstrated that electrospun nanofibres facilitated the adhesion of chondrocytes and helped to maintain smaller projected cell areas and a rounded cell phenotype, when compared to PLDLA films or microfibres. Random nanofibre composites were associated with the smallest and most rounded cells and aligned nanofibre composites also demonstrated a similar tendency. Quantitative PCR revealed that nanofibres promoted the expression of chondrogenic markers, such as collagen type IIaI and aggrecan, while maintaining low levels of collagen IaI. It was also found, by water contact angle measurement, that nanofibres were significantly more hydrophobic than cast films. The lower wettability of polymeric nanofibres favoured the maintenance of rounded chondrocyte morphology. To our knowledge this is the first study to confirm the positive influence on preserving chondrogenic phenotype and gene expression at the interface of true nano-microfibrous composites by using individual microfibres coated with aligned nanofibres. Such composites can potentially be fabricated into mechanically durable 3D scaffolds with better cell infiltration throughout the scaffolds.

Electrospinning: methods and development of biodegradable nanofibres for drug release.

It is clear that nanofibrous structures can be used as tools for many applications. It is already known that electrospinning is a highly versatile method of producing nanofibres and recent developments in the technique of electrospinning have led to the development of aligned nanofibres and biphasic, core-sheath fibres which can be used to encapsulate different materials from molecules to cells. Natural extracellular matrix (ECM) contains fibres in both micro and nano-scales and provides a structural scaffold which allows cells to localize, migrate, proliferate and differentiate. Polymer nanofibres can provide the structural cues of ECM. However, current literature gives new hope to further functionalising polymeric nanofibres by using them for drug delivery devices and improving their design to improve control of delivery. By encapsulating active agents within nanofibres (multifunctional nanofibres), a degree of control can be exerted over the release of encapsulated agents and therefore, the behaviour of cells can be manipulated for developing effective therapies and is extremely encouraging in the tissue engineering field by combining factors like fibre diameter, alignment and chemicals in new ways. Such multifunctional nanofibre-based systems are already being investigated in vivo. Experiments have shown the significant potential for treatments of disease and engineering of neural and bone tissues. Further, phase III clinical trials of nanofibrous patches for applications in wound treatment were encouraging. Hopefully, clinical applications of these drug delivery devices will follow, to enhance regenerative medicine applications.

Immunomodulatory effect of mesenchymal stromal cells: possible mechanisms.

Co-transplantation of mesenchymal stem cells (MSCs) and hematopoietic stem cells ameliorate hematopoietic reconstitution and induce tolerance. The immunomodulatory properties of MSCs have been demonstrated both in vivo and in vitro. MSCs can modulate function of immune cells such as T lymphocytes, antigen-presenting cells and natural killer cells. However, it is unknown whether MSCs given to patients that have undergone HSC transplantation could alleviate graft versus leukemia effect or could increase the risk of the infection. Proper characterization of MSC immunomodulatory mechanisms are crucial to anticipate the possible effect of MSC in the host. In the current report, interesting and contradictory results in the literature are reviewed in an attempt to understand the underlying mechanism. Differences in experimental designs and models used seem to be the underlying causes of discrepancy in reported results. Results of the few in vivo studies are controversial and further clinical studies are needed to confirm the efficiency and safety of MSCs in transplantation management.

Development of a bioactive glass fiber reinforced starch-polycaprolactone composite.

For bone regeneration and repair, combinations of different materials are often needed. Biodegradable polymers are often combined with osteoconductive materials, such as bioactive glass (BaG), which can also improve the mechanical properties of the composite. The aim of this work was to develop and characterize BaG fiber reinforced starch-poly-epsilon-caprolactone (SPCL) composite. Sheets of SPCL (30/70 wt %) were produced using single-screw extrusion. They were then cut and compression-molded in layers with BaG fibers to form composite structures with different combinations. Mechanical and degradation properties of the composites were studied. The actual amount of BaG in the composites was determined using combustion tests. Initial mechanical properties of the reinforced composites were at least 50% better than the properties of the nonreinforced specimens. However, the mechanical properties of the composites after 2 weeks of hydrolysis were comparable to those of the nonreinforced samples. During the 6 weeks hydrolysis the mass of the composites had decreased only by about 5%. The amount of glass in the composites remained as initial for the 6-week period of hydrolysis. In conclusion, it is possible to enhance initial mechanical properties of SPCL by reinforcing it with BaG fibers. However, mechanical properties of the composites are typical for bone fillers and strength properties need to be further improved for allowing more demanding bone applications.

Libyan National Health Services The Need to Move to Management-by-Objectives.

In the last four decades, there has been a substantial horizontal expansion of health services in Libya. This resulted in improvement in morbidity and mortality, in particularly those related to infectious disease. However, measures such as the national performance gap indicator reveal an underperforming health system. In this article, we discuss aspects related to the Libyan health system and its current status including areas of weakness. Overcoming current failures and further improvement are unlikely to occur spontaneously without proper planning. Defining community health problems, identifying unmet needs, surveying resources to meet them, establishing SMART (specific, measurable, achievable, and realistic and time specific) objectives, and projecting administrative action to accomplish the proposed programs, are a must. The health system should rely on newer approaches such as management-by-objectives and risk-management rather than the prevailing crisis-management attitude.

Potential use of craniosynostotic osteoprogenitors and bioactive scaffolds for bone engineering.

The cranial bone has a very limited regenerative capability. Patients with craniosynostosis (the premature fusion of cranial sutures, leading to skull abnormalities) often require extensive craniofacial reconstruction and repeated surgery. The possibility of grafting autologous osteoprogenitor cells seeded on bioabsorbable matrices is of great potential for inducing regeneration of craniofacial structure and protecting the brain from external insult. To this purpose we have studied the behaviour of normal and craniosynostotic mouse osteoblast cell lines, and of human primary osteoprogenitors from craniosynostotic patients. We have monitored their ability to grow and differentiate on plastic and on a scaffold composed of bioactive glass and bioabsorbable polymer by live fluorescent labelling and expression of bone differentiation markers. Cells from syndromic patients display a behaviour very similar to that observed in the stable mouse cell line we generated by introducing the human FGFR2-C278F, a mutation found in certain craniosynostosis, into MC3T3 osteblastic cells, indicating that the mutated cell line is a valuable model for studying the cellular response of human craniosynostotic osteoblasts. Both normal and mutated calvarial osteoprogenitors can attach to the bioactive scaffold, although mutated cells display adhesion defects when cultured on plastic. Furthermore, analysis of bone differentiation markers in human osteoblasts shows that the composite mesh, unlike PLGA(80) plates, supports bone differentiation. The ability of the mesh to support homing and differentiation in both normal and mutant osteoprogenitors is important, in view of further developing autologous biohybrids to repair cranial bone deficits also in craniosynostotic patients undergoing extensive reconstructive surgery.

Successful treatment of refractory tibial nonunion using calcium sulphate and bone marrow stromal cell implantation.

Successful healing of a nine-year tibial nonunion resistant to six previous surgical procedures was achieved by tissue engineering. We used autologous bone marrow stromal cells (BMSCs) expanded to 5 x 10(6) cells after three weeks' tissue culture. Calcium sulphate (CaSO4) in pellet form was combined with these cells at operation. The nonunion was clinically and radiologically healed two months after implantation. This is the description of on healing of a long-standing tibial nonunion by tissue engineering. The successful combination of BMSCs and CaSO4 has not to our knowledge been reported in a clinical setting.

Biodegradable nanomats produced by electrospinning: expanding multifunctionality and potential for tissue engineering.

With increasing interest in nanotechnology, development of nanofibers (n-fibers) by using the technique of electrospinning is gaining new momentum. Among important potential applications of n-fiber-based structures, scaffolds for tissue-engineering represent an advancing front. Nanoscaffolds (n-scaffolds) are closer to natural extracellular matrix (ECM) and its nanoscale fibrous structure. Although the technique of electrospinning is relatively old, various improvements have been made in the last decades to explore the spinning of submicron fibers from biodegradable polymers and to develop also multifunctional drug-releasing and bioactive scaffolds. Various factors can affect the properties of resulting nanostructures that can be classified into three main categories, namely: (1) Substrate related, (2) Apparatus related, and (3) Environment related factors. Developed n-scaffolds were tested for their cytocompatibility using different cell models and were seeded with cells for to develop tissue engineering constructs. Most importantly, studies have looked at the potential of using n-scaffolds for the development of blood vessels. There is a large area ahead for further applications and development of the field. For instance, multifunctional scaffolds that can be used as controlled delivery system do have a potential and have yet to be investigated for engineering of various tissues. So far, in vivo data on n-scaffolds are scarce, but in future reports are expected to emerge. With the convergence of the fields of nanotechnology, drug release and tissue engineering, new solutions could be found for the current limitations of tissue engineering scaffolds, which may enhance their functionality upon in vivo implantation. In this paper electrospinning process, factors affecting it, used polymers, developed n-scaffolds and their characterization are reviewed with focus on application in tissue engineering.

Novel drug delivery systems in pain therapy.

Pain is an unpleasant sensory experience resulting from damage to bodily tissues. It is considered a significant public health problem because it affects 1/5 of the world population and causes loss of great amounts of money. Pain reflects a mixture of pathological, psychological and genetic conditions that need deep understanding to be efficiently treated. If under-treated, pain results in serious immune and metabolic problems. Pain management faces many problems that limit its control. For instance, efficiency of pain killers is limited, pain killers give rise to serious side effects and inability of drug administration methods to help in pain control. Technology can overcome some of these problems and the introduction of implantable controlled drug delivery systems (CDDS), manufactured from biodegradable materials, offers a solution. Implantable CDDS provide good level of pain control, as they continuously provide drug, reduce side effects and improve patients' compliance. Biodegradable type of implantable CDDS are polymer based devices that are fabricated to locally deliver drugs in a pre-designed manner. They are currently a focus of research in the field of pain therapy in order to explore their chance to offer an alternative to the conventional methods for drug delivery. This paper aims to highlight the dimensions of pain issue and to overview the basics of drug release from polymers used for CDDS in pain management. In addition, it discusses the recent advances in the technologically designed drug delivery systems in the field of pain medicine and their clinical applications. Future perspectives are also presented.

PubMed Medical publications from Libya.

UNLABELLED: Medical research and publications are the back-bone for advancing the medical field. We identified the Pubmed medical publications that are affiliated with Libya to shed some light on the contribution of this country's medical community to the PubMed database. All publications affiliated with Libya in the PubMed were counted over a five year period ending December 2006. We also used the same method to obtain data on the PubMed medical publications from Tunisia, Morocco and Yemen. Tunisia had the largest number of PubMed publications among the studied countries: 20.4 publications per million population per year and 7.2 publications per year per one billion US$ GDP. Libya had much fewer publications: 2.4 publications per million population per year and 0.4 publications per one billion US$ GDP. The citation frequency for Libyan published research was very low compared to Tunisian and Moroccan related research. CONCLUSION: This preliminary analysis shows that medical research output in Libya is about twenty times less than in other countries with similar backgrounds, and that it needs to be enhanced.

PubMed Medical Publications From Libya

Medical research and publications are the back-bone for advancing the medical field. We identified the PubMed medical publications that are affiliated with Libya to shed some light on the contribution of this country's medical community to the PubMed database. All publications affiliated with Libya in the PubMed were counted over a five year period ending December 2006. We also used the same method to obtain data on the PubMed medical publications from Tunisia; Morocco and Yemen. Tunisia had the largest number of PubMed publications among the studied countries: 20.4 publications per million population per year and 7.2 publications per year per one billion US$ GDP. Libya had much fewer publications: 2.4 publications per million population per year and 0.4 publications per one billion US$ GDP. The citation frequency for Libyan published research was very low compared to Tunisian and Moroccan related research. Conclusion: This preliminary analysis shows that medical research output in Libya is about twenty times less than in other countries with similar backgrounds; and that it needs to be enhanced

Innovation in multifunctional bioabsorbable osteoconductive drug-releasing hard tissue fixation devices.

We review in this paper the work performed by our group to develop multifunctional bioabsorbable ciprofloxacin releasing bone implants. Poly lactide-co-glycolide (PLGA 80/20 and polylactide (P(L/DL)LA 70/30) were used. Ciprofloxacin (CF) and bioactive glass (BaG) 13-93 were added. The mixture was then extruded and self-reinforced. CF release, mechanical strength, and the effect on S. epidermidis attachment and biofilm formation were evaluated. In rabbits, tissue reactions were assessed. Pull out strength was evaluated in cadaver bones. CF was released over 44 weeks (P(L/DL)LA) and 23-26 weeks (PLGA). Initial shear strength of the CF screws was 152 MPa (P(L/DL)LA) and 172 MPa (PLGA). Strength was retained for 12 weeks (P(L/DL)LA) and 9 weeks (PLGA). Histologically, CF releasing implants did not show much difference from control plain PLGA screws except for increased giant cells. CF miniscrews had lower pullout strength than the controls, but CF tacks had better values than controls. BaG led to a drop in pullout strength properties. Bacterial growth, attachment and biofilm formation on CF implants was significantly reduced when compared to controls. Accordingly, bioabsorbable multifunctional implants with appropriate CF release, mechanical, and biocompatibility properties are possible to develop and are considered appropriate to apply clinically.

Electrospun multifunctional diclofenac sodium releasing nanoscaffold.

Electrospinning is a method utilized to produce nano-scale fibers for tissue engineering applications. A variety of cells are attracted by nano scale surfaces and structures probably due to the similarity of their natural environment scale. In this study, diclofenac sodium (DS) releasing nanofibers were manufactured via electrospinning process. Poly(95 epsilon-capro/5 D,L-lactide) was dissolved into acetic acid to form a 20% w/v solution. 2% w/w of DS was then added into the polymer solution and stirred homogenously. About 1 g of polymer/drug solution was spun onto the collector under electrostatic conditions. The distance between needle tip and sample collector was arranged to 10 cm and applied electric field was 2 kV/cm. Release rate of DS was measured by using UV/VIS spectrophotometer. Resulted highly porous nanofiber scaffold was about 2 mm thick and the diameter of nanofibers was approximately 130 nm. Structure included in also spheres with approximately diameter of 3.30 microm. About 45% of DS was released during the first 24 hours and after that the release decreased to almost zero value. After 35 days release rate increased. This study revealed that manufacturing of highly porous DS releasing nanoscaffold by electrospinning process is feasible. Having fast DS release rate nanofibrous scaffold made of poly(95 epsilon-capro/5 D,L-lactide) can be of benefit for applications where immediate control of tissue reaction is needed.

Development of diclofenac sodium releasing bio-erodible polymeric nanomats.

Application of nanofiber-based nanomats in medicine is attractive and thanks to the 3D nanostructure and the high surface to volume ratio they are excellent for local controlled drug delivery. The use of bioactive bioerodible polymers for developing drug delivery nanomats may allow for drug release and targeting control. Objective of the current study was to evaluate the suitability of bioerodible polymeric material based on n-butyl hemiester of [poly(maleic anhydride-alt-2-methoxyethyl vinyl ether)] (PAM14) for the preparation of nanomats for controlled administration of anti-inflammatory, diclofenac sodium (DS) drug. Samples were prepared using different polymer concentrations (5-10%) in either ethanol or acetic acid as solvent. Morphology was investigated by using scanning electron microscopy (SEM). Thermal analysis such as differential scanning calorimetry (DSC) was performed to detect effect on polymer arrangement. DS localization in electrospun nanomats was evaluated by using electron back scattering microanalysis, based on the detection of chlorine, and drug release kinetics was assessed using UV-Vis. Average fiber diameter resulted in the range of 100 nm to 1.0 microm and a homogeneous distribution of the loaded drug into the fibers was observed. The DS release was immediate and despite the preliminary nature of the performed electrospinning experiments, the achieved results appear promising for the future development of a novel system for the controlled and targeted administration of drug and active agent.

Histomorphometric analysis of poly-L/D-lactide 96/4 sutures in the gastrocnemius tendon of rabbits.

BACKGROUND: Common Achilles tendon ruptures are not usually fixed by bioabsorbable sutures due to limitations in their strength retention properties. Modern technology has made it possible to develop bioabsorbable sutures with prolonged strength retention. AIMS: To evaluate histologically tissue reactions of poly-L/D-lactide (PLDLA) sutures implanted in Achilles tendon of rabbits. MATERIAL AND METHODS: Fifteen rabbits were evaluated at 2, 6 and 12 weeks postoperatively, with five rabbits in each follow-up group. PLDLA monofilament sutures were implanted into the medial gastrocnemius tendon. Polyglyconate monofilament sutures with similar diameter (Maxon 4-0, Cyanamid of Great Britain Ltd., Gosport, UK) were implanted in the contralateral gastrocnemius tendon. The histology was studied in hard-resin embedded samples. The thickness of the formed fibrous tissue capsule was determined histomorphometrically. RESULTS: PLDLA led to formation of significantly thinner fibrous tissue capsule than Maxon sutures of the same diameter. Median thickness (PLDLA vs. Maxon) at two weeks was 5.26 vs.13.22 microm, at six weeks 11.66 vs. 80.97 microm, and at 12 weeks 10.63 vs. 17.59 microm (p<0.01). CONCLUSIONS: During the 12 week follow-up period, PLDLA sutures implanted intratendineously formed thinner fibrous capsule than Maxon sutures of the same diameter. The suture materials were not totally absorbed by 12 weeks.

Biodegradable nanomats produced by electrospinning: expanding multifunctionality and potential for tissue engineering.

With increasing interest in nanotechnology, development of nanofibers (n-fibers) by using the technique of electrospinning is gaining new momentum. Among important potential applications of n-fiber-based structures, scaffolds for tissue-engineering represent an advancing front. Nanoscaffolds (n-scaffolds) are closer to natural extracellular matrix (ECM) and its nanoscale fibrous structure. Although the technique of electrospinning is relatively old, various improvements have been made in the last decades to explore the spinning of submicron fibers from biodegradable polymers and to develop also multifunctional drug-releasing and bioactive scaffolds. Various factors can affect the properties of resulting nanostructures that can be classified into three main categories, namely: (1) Substrate related, (2) Apparatus related, and (3) Environment related factors. Developed n-scaffolds were tested for their cytocompatibility using different cell models and were seeded with cells for to develop tissue engineering constructs. Most importantly, studies have looked at the potential of using n-scaffolds for the development of blood vessels. There is a large area ahead for further applications and development of the field. For instance, multifunctional scaffolds that can be used as controlled delivery system do have a potential and have yet to be investigated for engineering of various tissues. So far, in vivo data on n-scaffolds are scarce, but in future reports are expected to emerge. With the convergence of the fields of nanotechnology, drug release and tissue engineering, new solutions could be found for the current limitations of tissue engineering scaffolds, which may enhance their functionality upon in vivo implantation. In this paper electrospinning process, factors affecting it, used polymers, developed n-scaffolds and their characterization are reviewed with focus on application in tissue engineering.