Urologic Robotic Surgery.

Urologists have always been leaders in advancing surgical technology and were the first to utilize modern robotic surgery for robotic-assisted laparoscopic radical proctectomy. Surgeon ergonomics, instrument precision, operative time, and postoperative recovery were all objectively improved. In urology, robotic surgery is now used for all intra-abdominal, retroperitoneal, and pelvic procedures and has been expanded to renal transplants and pediatric use. Modern robotic surgery has become an essential part of treating complex urologic disease in the developed world. Urologists continue to lead the way with the latest robotic surgical systems, including the newly approved single port systems.

Remote-controlled internal lengthening plate for distraction osteogenesis in pediatric patients.

OBJECTIVES: Limb lengthening by distraction osteogenesis is a technique widely used to treat limb length discrepancy resulting from trauma, congenital limb defects and long bone non-union. For decades, patients have resorted to the Ilizarov apparatus, prone to pin tract infections and scarring. Although implantable lengthening nails have reduced the incidence of complications, they are not applicable in pediatric patients with open growth plates. The aim of this project is to design a remote-controlled internal lengthening device suitable for implantation in children. METHODS: The proposed device has the form of an internal remote-controlled telescopic lengthening plate, screwed to the lateral side of the bone with locking screws. This is appropriate for use with pediatric patients. It has been tested on an experimental bench which has the form of a vertically sliding platform, on which were stacked weights simulating soft-tissue resistance forces. RESULTS: This internal lengthening plate generated distraction forces of up to 735 N on wooden and synthetic bones (SawbonesTM). Furthermore, it maintained a constant distraction speed over the course of the procedure for a given weight. CONCLUSIONS: This device represents a major advancement in the field of pediatric limb-lengthening, addressing a demographic gap left open by current implantable devices.

One-step fabrication of apatite-chitosan scaffold as a potential injectable construct for bone tissue engineering.

Biomineralization of soft scaffolds is a new venture in bone tissue engineering. This work aimed to develop a new injectable and in-situ gelling soft scaffold with chitosan and apatites through a rapid purine-crosslinking reaction. The scaffolds were fabricated by mixing chitosan, adenosine diphosphate and biominerals in an 'all-in-one-step' procedure. The gelling of chitosan via the crosslinker occurs in <4 s as measured by impedance spectroscopy. These soft gels could retain up to 4 times their weight in water. Spectroscopy showed the formation of ionic bonds between chitosan and the apatites. Morphological analyses revealed an interconnected, highly porous structure, with pore size ranging from 200 nm-200 µm that was maintained even with the biominerals. Rheology showed a viscoelastic behavior of the solutions and the elastic behavior of the sponges, and therefore their injectability potential. in vitro studies showed good cell adhesion and morphology, as well as potential use for bone tissue engineering applications.

The bioconjugation mechanism of purine cross-linkers affects microstructure and cell response to ultra rapidly gelling purine-chitosan sponges.

As a cell carrier, cross-linking is one of the most common approaches used to provide chitosan with greater structural integrity. We introduced a cross-linking strategy by using two purines, guanosine 5'-diphosphate (GDP) or adenosine 5'-diphosphate (ADP), as cross-linkers. The rationale for this approach is that both GDP and ADP have an important physiological role and act as intercellular signaling molecules in numerous biological processes. The slight difference between the chemical structure of guanine and adenosine in GDP and ADP, respectively, affect the cross-linking mechanism. This resulted in a different scaffold microstructure and thus, altered the response of encapsulated cells to the scaffold. FTIR and solid-state 13C-NMR revealed the formation of a quadruplex structure among the four GDP molecules confined between the chitosan backbone. This resulted from the ability of guanine to form intermolecular hydrogen bonds, while adenosine in ADP lacks this capacity. The formation of a more organized structure in GDP-chitosan sponges also increased the crystallinity of the sponge as shown by X-ray diffraction data. Further, physicochemical analyses with SEM and µCT indicated a more open-pore architecture and increased porosity. Although an active population of encapsulated cells was maintained in all chitosan sponges overtime, the GDP-based sponges provided a 6-fold increase in the activity of MC-3T3 cells and significantly enhanced their proliferation due to a more appropriate microstructure. Overall, these findings suggest that slight changes in the chemical structure of the cross-linker in the preparation of chitosan-based biomaterials will have a significant impact on the structural properties of the chitosan. This important parameter can be utilized to modulate cell response and to understand the cell signaling pathway of chitosan-based biomaterials in the context of their applications in tissue engineering.

Small Players Ruling the Hard Game: siRNA in Bone Regeneration.

Silencing gene expression through a sequence-specific manner can be achieved by small interfering RNAs (siRNAs). The discovery of this process has opened the doors to the development of siRNA therapeutics. Although several preclinical and clinical studies have shown great promise in the treatment of neurological disorders, cancers, dominant disorders, and viral infections with siRNA, siRNA therapy is still gaining ground in musculoskeletal tissue repair and bone regeneration. Here we present a comprehensive review of the literature to summarize different siRNA delivery strategies utilized to enhance bone regeneration. With advancement in understanding the targetable biological pathways involved in bone regeneration and also the rapid progress in siRNA technologies, application of siRNA for bone regeneration has great therapeutic potential. High rates of musculoskeletal injuries and diseases, and their inevitable consequences, impose a huge financial burden on individuals and healthcare systems worldwide.

A combinatorial approach towards achieving an injectable, self-contained, phosphate-releasing scaffold for promoting biomineralization in critical size bone defects.

An injectable, guanosine 5'-diphosphate (GDP)-crosslinked chitosan sponge was investigated as a drug delivery system (DDS) for accelerating biomineralization in critical size bone defects (CSBDs). Two approaches were examined both individually, and in combination, in order to achieve this goal. The first approach involved the encapsulation and release of Bone Morphogenetic Protein 7 (BMP-7), a powerful mineralization stimulant. Results confirmed that the rapid gelation of the chitosan sponge prompted high encapsulation of BMP-7 and provided a controlled release over a period of 30 days with no burst release. The second approach was aimed at encapsulating pyrophosphatase (PPtase) in the chitosan sponge to cleave pyrophosphate (PPi) - a mineralization inhibitor and a degradation by-product of the chitosan sponge - into phosphate ions (Pi). PPtase was successfully encapsulated in the chitosan sponge and was able to completely eliminate PPi from the media by cleaving them to Pi. Chitosan sponges releasing Pi into the media were shown to increase overall biomineralization fourfold as compared to controls, an amount equivalent to biomineralization caused by direct injection of 1µg of free BMP-7 to the cells. Even though the combined encapsulation of 1µg BMP-7 and PPtase in the sponges did not demonstrate an additional increase in biomineralization, encapsulation of low concentrations of BMP-7 can promote mesenchymal stem cell migration into the sponge after application in vivo. The findings suggest that the sponge-PPtase system likely allows excellent bone regeneration with lower concentrations of BMP-7, reducing risks and expense of the treatment. STATEMENT OF SIGNIFICANCE: There are bone defects, known as critical size defects, which do not heal on their own and require a therapeutic intervention. The current commercially-available therapies use large quantities of growth factors, such as Bone Morphogenetic Proteins (BMPs), which makes them expensive and a source for a myriad of unwanted side effects. In this manuscript we demonstrate, for the first time, the use of an injectable chitosan-based sponge that contains no inorganic components, but can nonetheless act as a source of phosphate ions to improve bone mineralization. We also demonstrate that this sponge can entrap small concentrations of BMP-7 and provide controlled release over time. The ability to release phosphate ions and low concentrations of BMP-7 makes this therapeutic intervention clinically-relevant, affordable, and safe.

Purine-crosslinked injectable chitosan sponges promote oligodendrocyte progenitor cells' attachment and differentiation.

Oligodendrocyte Progenitor Cells (OPCs) reside in the central nervous system (CNS) and are responsible for remyelinating axons after a spinal cord injury (SCI). However, the remyelination process is incomplete and abnormal due to the inability of OPCs to fully differentiate at the site of injury. In this study a newly developed injectable chitosan sponge crosslinked using guanosine 5'-diphosphate (GDP) was used to enhance OPC survival, attachment and differentiation. This purine-based biomaterial is the first of its kind and its inception was based on the growing body of literature concerning the role of purinergic signalling in the CNS. GDP-crosslinked chitosan sponges are rapidly-gelling and can be easily administered in situ using an injection system based on a double-lumen design. The chitosan sponges prompted OPC differentiation even in the presence of mitogens. Moreover, neurotrophin-3 (NT-3) was successfully entrapped in the sponges and a sustained release for up to 30 days was achieved. OPCs were shown to differentiate into mature oligodendrocytes that express myelin basic protein (MBP) when cultured on sponges containing NT-3. These findings, along with the suitable physicochemical and biological properties, make these sponges conducive to use as viable therapeutic agents for enhancing remyelination post-SCI.

Genipin-crosslinked chitosan/poly-L-lysine gels promote fibroblast adhesion and proliferation.

Chitosan blends have been widely investigated to create biomaterials with desirable physicochemical and biological properties for tissue engineering applications. A recurring difficulty, however, has been to maintain their stability in an aqueous environment. The rationale behind this study was to demonstrate that genipin crosslinking can improve and maintain the stability of chitosan/poly-l-lysine (PLL) blends. Four gel formulations were prepared by varying the weight ratios of chitosan and PLL. Electron microscopy revealed that genipin crosslinking provided a more homogenous gel surface compared to uncrosslinked gels. Moreover, it was discovered that 3h was sufficient to stabilize the gels. In vitro studies using fibroblasts demonstrated that genipin-crosslinked gels enhanced fibroblasts' attachment as compared to uncrosslinked gels. Moreover, cell viability was significantly improved by 1.6 times on 60:40 gels, and 6.5 times on 50:50 gels after crosslinking. Finally, proliferation was enhanced up to 5 times on 60:40 gels.

Injectable chitosan-based scaffolds in regenerative medicine and their clinical translatability.

Injectable scaffolds (IS) are polymeric solutions that are injected in vivo and undergo gelation in response to physiological or non-physiological stimuli. Interest in using IS in regenerative medicine has been increasing this past decade. IS are administered in vivo using minimally invasive surgery, which reduces hospitalization time and risk of surgical wound infection. Here, chitosan is explored as an excellent candidate for developing IS. A literature search reveals that 27% of IS publications in the past decade investigated injectable chitosan scaffolds (ICS). This increasing interest in chitosan stems from its many desirable physicochemical properties. The first section of this Progress Report is a comprehensive study of all physical, chemical, and biological stimuli that have been explored to induce ICS gelation in vivo. Second, the use of ICS is investigated in four major regenerative medicine applications, namely bone, cartilage, cardiovascular, and neural regeneration. Finally, an overall critique of the ICS literature in light of clinical translatability is presented. Even though ICS have been widely explored in the literature, very few have progressed to clinical trials. The authors discuss the current barriers to moving ICS into the clinic and provide suggestions regarding what is needed to overcome those challenges.

Rapid, guanosine 5'-diphosphate-induced, gelation of chitosan sponges as novel injectable scaffolds for soft tissue engineering and drug delivery applications.

Novel injectable chitosan sponges based on rapid ionic crosslinking using guanosine 5'-diphosphate are introduced. The rapid gelation, high water retention, desirable physicochemical properties, soft tissue-like mechanical properties, and excellent cytocompatibility make these injectable sponges promising candidates for tissue regeneration and drug delivery applications.

Oligodendrocyte-protection and remyelination post-spinal cord injuries: a review.

In the past four decades, the main focus of investigators in the field of spinal cord regeneration has been to devise therapeutic measures that enhance neural regeneration. More recently, emphasis has been placed on enhancing remyelination and providing oligodendrocyte-protection after a spinal cord injury (SCI). Demyelination post-SCI is part of the cascading secondary injury that takes place immediately after the primary insult; therefore, therapeutic measures are needed to reduce oligodendrocyte death and/or enhance remyelination during the acute stage, preserving neurological functions that would be lost otherwise. In this review a thorough investigation of the oligodendrocyte-protective and remyelinative molecular therapies available to date is provided. The advent of new biomaterials shown to promote remyelination post-SCI is discussed mainly in the context of a combinatorial approach where the biomaterial also provides drug delivery capabilities. The aim of these molecular and biomaterial-based therapies is twofold: (1) oligodendrocyte-protective therapy, which involves protecting already existing oligodendrocytes from undergoing apoptosis/necrosis; and (2) inductive remyelination, which involves harnessing the remyelinative capabilities of endogenous oligodendrocyte precursor cells (OPCs) at the lesion site by providing a suitable environment for their migration, survival, proliferation and differentiation. From the evidence reported in the literature, we conclude that the use of a combinatorial approach including biomaterials and molecular therapies would provide advantages such as: (1) sustained release of the therapeutic molecule, (2) local delivery at the lesion site, and (3) an environment at the site of injury that promotes OPC migration, differentiation and remyelination.

Genipin-cross-linked electrospun collagen fibers.

The fabrication of a fibrous collagen scaffold using electrospinning is desirable for tissue-engineering applications. Previously, electrospun collagen fibers were shown to be unstable in aqueous environments and, therefore, cross-linking is essential to stabilize these fibers. In this study genipin, a significantly less cytotoxic cross-linking agent compared to glutaraldehyde, was used to cross-link electrospun collagen fibers. The significance of this research lies in the use of four alcohol/water solvent systems to carry out the crosslinking reaction to maintain fibrous morphology during cross-linking. The four cross-linking conditions established were: (1) ethanol, 5% water and 3 days, (2) ethanol, 3% water and 5 days, (3) ethanol, 5% water and 5 days, and (4) isopropanol, 5% water and 5 days at a genipin concentration of 0.03 M. Results illustrated that genipin-cross-linking was effective in maintaining collagen fiber integrity in aqueous and cell culture media environments for up to 7 days. In addition, it was shown that fiber swelling could be controlled by using different cross-linking conditions. Swelling of cross-linked fibers immersed in Dulbecco's modified eagle medium for 7 days ranged from 0 to 59 ± 4%. The cross-linked fibers were analyzed using scanning electron microscopy, Fourier transform infrared spectroscopy and ninhydrin assay. Finally, studies using primary human fibroblasts indicated good cell adhesion to these scaffolds. Overall, our data suggest that these stabilized fibrous collagen scaffolds provide a promising environment for tissue-regeneration applications.