RESUMO
Many factors contribute to the poor axonal regrowth and ineffective functional recovery after spinal cord injury (SCI). Biomaterials have been used for SCI repair by promoting bridge formation and reconnecting the neural tissue at the lesion site. The mechanical properties of biomaterials are critical for successful design to ensure the stable support as soon as possible when compressed by the surrounding spine and musculature. Poly(propylene fumarate) (PPF) scaffolds with high mechanical strength have been shown to provide firm spatial maintenance and to promote repair of tissue defects. A multichannel PPF scaffold is combined with collagen biomaterial to build a novel biocompatible delivery system coated with neurotrophin-3 containing an engineered collagen-binding domain (CBD-NT3). The parallel-aligned multichannel structure of PPF scaffolds guide the direction of neural tissue regeneration across the lesion site and promote reestablishment of bridge connectivity. The combinatorial treatment consisting of PPF and collagen loaded with CBD-NT3 improves the inhibitory microenvironment, facilitates axonal and neuronal regeneration, survival of various types of functional neurons and remyelination and synapse formation of regenerated axons following SCI. This novel treatment strategy for SCI repair effectively promotes neural tissue regeneration after transected spinal injury by providing a regrowth-supportive microenvironment and eventually induces functional improvement.
Assuntos
Fumaratos/química , Fatores de Crescimento Neural/química , Regeneração Nervosa/fisiologia , Polipropilenos/química , Traumatismos da Medula Espinal/terapia , Alicerces Teciduais/química , Animais , Ratos , Engenharia TecidualRESUMO
Therapeutic stem cell transplantation bears the promise of new directions in organ and tissue replacement, but a number of its difficulties and perils are also well known. Our goal was to develop a method of transplantation by which the transplanted cells remain confined to the transplantation site and induce favorable processes. With the help of mask-projection excimer laser stereolithography, 3D hybrid nanoscaffolds were fabricated from biodegradable, photocurable PPF:DEF resin with incorporated gold nanoparticles (Au NPs). The scaffolds were tested in vitro and in vivo in order to find out about their biocompatibility and fitness for our purposes. In vitro, macrophages and mouse autologous adipose stem cells (ASCs) were seeded over the hybrid scaffolds and non-hybrid (with Au NPs) scaffolds for 4days. The hybrid nanocomposite greater stem cell dispension and stem cell adhesion than PPF scaffolds without Au NPs, but such a difference was not seen in the case of macrophages. In vivo, stem cells, scaffoldings and scaffoldings covered in stem cells were transplanted under the back skin of mice. After 14days, blood samples were taken and the affected skin area was excised. Cytokine and chemokine profiling did not indicate elevated immunomediators in the sera of experimental animals. Interestingly, the autologous-stem-cell-seeded hybrid nanocomposite scaffold induced muscle tissue regeneration after experimental wound generation in vivo. We could not observe such stem cell-induced tissue regeneration when no scaffolding was used. We conclude that PPF:DEF resin nanoscaffolds with incorporated gold nanoparticles offer a safe and efficient alternative for the enhancement of local tissue remodeling. The results also support the idea that adipose derived stem cells are an optimal cell type for the purposes of regenerative musculoskeletal tissue engineering.
Assuntos
Materiais Biocompatíveis/química , Ouro/química , Nanopartículas Metálicas/química , Polímeros/química , Alicerces Teciduais/química , Tecido Adiposo/citologia , Animais , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Quimiocinas/metabolismo , Citocinas/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Pele/patologia , Transplante de Células-Tronco , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Engenharia TecidualRESUMO
The effects of various fabrication parameters of our Mask Projection Excimer Laser StereoLithography (MPExSL) system were investigated. We demonstrate that laser parameters directly change the physical properties (stiffness, thermal degradation, and height/thickness) of the poly(propylene fumarate) (PFF) scaffold structures. The tested parameters were the number of pulses, fluence per pulse and laser repetition rate. We present a four-order tuning capability of MPExSL-fabricated structures' stiffness without altering the resin composition or using cumbersome post-treatment procedures. Thermogravimetric analysis and differential scanning calorimetry confirmed this tuning capability. Prototype-segmented scaffold designs are presented and analyzed to further expand the concept and exploit this in situ stiffness tuning capability of the scaffolds for tissue engineering and regenerative medicine applications.
Assuntos
Materiais Biocompatíveis/química , Polímeros/química , Alicerces Teciduais/química , Fumaratos/química , Lasers , Teste de Materiais/métodos , Polipropilenos/química , Porosidade , Engenharia Tecidual/métodosRESUMO
Fibers of cellulose networks were stably coated with N-methacrylate glycol chitosan (MGC) shells using subsequent steps of dip coating and photo-curing. The photo-crosslinked MGC-coated cellulose networks preserved their fibrous structure. A model hydrophilic antiseptic solution containing eosin, chloroxylenol and propylene glycol was incorporated into the shells to study the drug release dynamics. Detailed drug release mechanism into phosphate buffered saline (PBS) solutions from coated and pristine fibers loaded with the antiseptic was investigated. The results show that the MGC-coated cellulose fibers enable the controlled gradual release of the drug for four days, as opposed to fast, instantaneous release from eosin coated pristine fibers. This release behavior was found to affect the antibacterial efficiency of the fibrous cellulose sheets significantly against Staphylococcus aureus and Candida albicans. In the case of the MGC-eosin functionalized system the antibacterial efficiency was as high as 85% and 90%, respectively, while for the eosin coated pristine cellulose system the efficiency was negative, indicating bacterial proliferation. Furthermore, the MGC-eosin system was shown to be efficacious in a model of wound healing in mice, reducing the levels of various pro-inflammatory cytokines that modulate early inflammatory phase responses. The results demonstrate good potential of these coated fibers for wound dressing and healing applications. Due to its easy application on common passive commercial fibrous dressings such as gauzes and cotton fibers, the method can render them active dressings in a cost effective way.
Assuntos
Anti-Infecciosos Locais/farmacologia , Celulose/química , Quitosana/química , Amarelo de Eosina-(YS)/farmacologia , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Animais , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Linhagem Celular , Celulose/ultraestrutura , Preparações de Ação Retardada , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pele/efeitos dos fármacos , Espectrometria por Raios X , Espectrofotometria UltravioletaRESUMO
One of the main open issues in modern vascular surgery is the nonbiodegradability of implants used for stent interventions, which can lead to small caliber-related thrombosis and neointimal hyperplasia. Some new, resorbable polymeric materials have been proposed to substitute traditional stainless-steel stents, but so far they were affected by poor mechanical properties and low biocompatibility. In this respect, a new material, polypropylene fumarate (PPF), may be considered as a promising candidate to implement the development of next generation stents, due to its complete biodegradability, and excellent mechanical properties and the ease to be precisely patterned. Besides all these benefits, PPF has not been tested yet for vascular prosthesis, mainly because it proved to be almost inert, while the ability to elicit a specific biological function would be of paramount importance in such critical surgery applications. Here, we propose a biomimetic functionalization process, aimed at obtaining specific bioactivation and thus improved cell-polymer interaction. Porous PPF-based scaffolds produced by deep-UV photocuring were coated by elastin and the functionalized scaffolds were extensively characterized, revealing a stable bound between the protein and the polymer surface. Both 3T3 and HUVEC cell lines were used for in vitro tests displaying an enhancement of cells adhesion and proliferation on the functionalized scaffolds.