RESUMO
Liquid, injectable hydrophobic polymers have advantages as degradable drug delivery vehicles; however, polymers examined for this purpose to date form acidic degradation products that may damage acid-sensitive drugs. Herein, we report on a new viscous liquid vehicle, poly(trimethylene carbonate-co-5-hydroxy-trimethylene carbonate), which degrades through intramolecular cyclization producing glycerol, carbon dioxide, and water-soluble trimethylene carbonate. Copolymer degradation durations from weeks to months were achieved with the 5-hydroxy-trimethylene carbonate (HTMC) content of the oligomer having the greatest impact on the degradation rate, with oligomers possessing a higher HTMC content degrading fastest. The degradation products were non-cytotoxic towards 3T3 fibroblasts and RAW 264.7 macrophages. These copolymers can be injected manually through standard gauge needles and, importantly, during in vitro degradation, the microenvironmental pH within the oligomers remained near neutral. Complete and sustained release of the acid-sensitive protein vascular endothelial growth factor was achieved, with the protein remaining highly bioactive throughout the release period. These copolymers represent a promising formulation for local and sustained release of acid sensitive drugs.
Assuntos
Carbonatos/química , Dioxanos/química , Preparações Farmacêuticas/química , Polímeros/química , Água/química , Células 3T3 , Animais , Dióxido de Carbono/química , Química Farmacêutica/métodos , Sistemas de Liberação de Medicamentos/métodos , Excipientes/química , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Concentração de Íons de Hidrogênio , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Células RAW 264.7 , Fator A de Crescimento do Endotélio Vascular/metabolismo , ViscosidadeRESUMO
Intravitreal sustained delivery of corticosteroids such as dexamethasone is an effective means of treating a number of ocular diseases, including diabetic retinopathy, uveitis, and age-related or diabetic macular edema. There are currently marketed devices for this purpose, yet only one, Ozurdex, is degradable. In vitro release of dexamethasone from the Ozurdex device is limited to approximately 30 days, however. It was the objective of this study to examine the potential for prolonged and sustained release of a corticosteroid in vitro from a degradable polymer prepared from terminally acrylated star co- and ter-prepolymers composed of d,l-lactide, ε-caprolactone, and trimethylene carbonate co-photo-cross-linked with poly(ethylene glycol) diacrylate. Through manipulation of the network polymer glass transition temperature and degradation rate, a sustained release of triamcinolone was achieved, with an estimated release duration greater than twice that of the Ozurdex system. Moreover, a period of nearly constant release was obtained using a network prepared from 5000 Da star-poly(trimethylene carbonate-co-d,l-lactide) triacrylate (3:1 trimethylene carbonate:d,l-lactide) co-cross-linked with 700 Da poly(ethylene glycol diacrylate). These formulations show promise as implantable, intravitreal corticosteroid delivery devices.
Assuntos
Corticosteroides/administração & dosagem , Corticosteroides/química , Injeções Intraoculares/métodos , Polímeros/química , Caproatos/química , Retinopatia Diabética , Sistemas de Liberação de Medicamentos/métodos , Lactonas/química , Triancinolona/químicaRESUMO
Melt electrospinning writing (MEW) is an emerging additive manufacturing technique that enables the design and fabrication of micrometer-thin fibrous scaffolds made of biocompatible and biodegradable polymers. By using a computer-aided deposition process, a unique control over pore size and interconnectivity of the resulting scaffolds is achieved, features highly interesting for tissue engineering applications. However, MEW has been mainly used to process low melting point thermoplastics such as poly(ε-caprolactone). Since this polymer exhibits creep and a reduction in modulus upon hydration, we manufactured scaffolds of poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) (poly(LLA-ε-CL-AC)), a photo-cross-linkable and biodegradable polymer, for the first time. We show that the stiffness of the scaffolds increases significantly (up to â¼10-fold) after cross-linking by UV irradiation at room temperature, compared with un-cross-linked microfiber scaffolds. The preservation of stiffness and high average fiber modulus (370 ± 166 MPa) within the cross-linked hydrated scaffolds upon repetitive loading (10% strain at 1 Hz up to 200,000 cycles) suggests that the prepared scaffolds may be of potential interest for soft connective tissue engineering applications. Moreover, the approach can be readily adapted through manipulation of polymer properties and scaffold geometry to prepare structures with mechanical properties suitable for other tissue engineering applications.
Assuntos
Materiais Biocompatíveis/química , Plásticos Biodegradáveis/química , Tecido Conjuntivo/fisiologia , Polímeros/síntese química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Poliésteres/química , Polímeros/química , Impressão TridimensionalRESUMO
The aim of this study was to prepare a fibrous scaffold that possesses a crimped morphology using a photo-cross-linkable biodegradable copolymer. To obtain the crimped morphology, the polymer was first electrospun onto a rotating wire mandrel to obtain aligned straight fibers. Postprocessing by immersion in aqueous buffer at 37 °C generated a crimplike pattern in the fibers. It was reasoned that cross-linking the fibers following formation of the crimped structure would endow the scaffolds with a recoverable crimp pattern and mechanical properties similar to that of the collagen fibers in the anterior cruciate ligament (ACL). To achieve this aim, a trimethylene carbonate based monomer bearing an acrylate pendant group was synthesized and copolymerized with l-lactide. The copolymer was electrospun and photo-cross-linked yielding fibrous scaffolds possessing a substantial increase in tensile modulus and crimp stability compared to the uncross-linked fibrous scaffolds. The crimp-stabilized scaffolds also showed good cytocompatibility toward 3T3 fibroblasts, which attached and grew along the crimped fibers. These findings suggest that these cross-linked fiber scaffolds may be useful for the generation of cultured ligament tissue.
Assuntos
Resinas Acrílicas/química , Materiais Biocompatíveis/química , Reagentes de Ligações Cruzadas/química , Poliésteres/química , Engenharia Tecidual , Alicerces Teciduais/química , Resinas Acrílicas/síntese química , Animais , Materiais Biocompatíveis/síntese química , Células Cultivadas , Reagentes de Ligações Cruzadas/síntese química , Fibroblastos/citologia , Camundongos , Estrutura Molecular , Células NIH 3T3 , Poliésteres/síntese químicaRESUMO
Thiol-modified nanoparticles have potential applications in mucoadhesive drug delivery and have been examined in this regard for topical ocular delivery. In this paper we provide a simple method for the synthesis of a dithiol terminated amphiphilic diblock copolymer. Bidentate dithiol-poly(ethylene glycol)-poly(d,l-lactide) (SH2-PEG-PDLLA) was synthesized and micelles with dithiol-containing coronas were prepared from this block copolymer via the emulsion method. In vitro release studies indicated that the presence of the thiol groups at the surface did not affect the rate of release of dexamethasone, used as a representative ocular drug. The micelles also showed low cytotoxicity to human corneal epithelial cells (HCEC) and murine fibroblast cells (3T3 cells). A hydrophobic red fluorophore, Nile red, was loaded into the core of micelles and confocal microscopy was used to study HCEC uptake and retention of the micelles. The micelles were rapidly endocytosed by the HCEC, with intracellular micelle levels remaining unchanged with incubation times from 5 to 120 min. Interestingly, Nile red was eliminated significantly more slowly from HCECs treated with the thiolated micelles. These results suggest that these dithiolated micelles may be effective for topical ocular drug delivery.
Assuntos
Administração Oftálmica , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Polímeros/síntese química , Células 3T3/efeitos dos fármacos , Animais , Células Cultivadas , Dexametasona/administração & dosagem , Dexametasona/farmacocinética , Portadores de Fármacos/síntese química , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Células Epiteliais/efeitos dos fármacos , Epitélio Corneano/citologia , Epitélio Corneano/efeitos dos fármacos , Corantes Fluorescentes/farmacocinética , Humanos , Camundongos , Micelas , Oxazinas/farmacocinética , Poliésteres/química , Polietilenoglicóis/química , Polímeros/química , Tolueno/análogos & derivados , Tolueno/químicaRESUMO
Intervertebral disk degeneration (IVDD) is a common condition that causes severe back pain and affects patients' mobility and life quality considerably. IVDD originates within the central region of the disk called the nucleus pulposus (NP). Removing the damaged tissue and replacing it with NP cells (NPCs) delivered within an in situ forming hydrogel is a promising treatment approach. Herein we describe a hydrogel formulation based on 4-arm [poly(ethylene glycol)-b-poly(trimethylene carbonate)-acrylate] (4a[PEG-b-PTMC-Ac]) crosslinked with thiolated chondroitin sulfate via Michael-type reaction for this purpose. A library of hydrogels based on 15 kDa 4a-[PEG] with PTMC blocks of varying molecular weight were prepared and characterized. The instantaneous moduli of the hydrogels were adjustable from 24 to 150 kPa depending on the length of the PTMC block and the polymer volume fraction. The influence of each of these parameters was effectively explained using both scaling or mean field theories of polyelectrolyte hydrogels. The hydrogels were resistant to cyclic compressive loading and degraded gradually over 70 days in vitro. A hydrogel formulation with an instantaneous modulus at the high end of the range of values reported for human NP tissue was chosen to assess the ability of these hydrogels for delivering NPCs. The prepolymer solution was injectable and formed a hydrogel within 30 minutes at 37 °C. Bovine NPCs were encapsulated within this hydrogel with high viability and proliferated throughout a 28 day, hypoxic culture period. The encapsulated NPCs formed clusters and deposited collagen type II but no collagen type I within the hydrogels. Despite an initial gradual decrease, a steady-state modulus was reached at the end of the 28 day culture period that was within the range reported for healthy human NP tissue. This in situ forming hydrogel formulation is a promising approach and with further development could be a viable clinical treatment for IVDD.
Assuntos
Degeneração do Disco Intervertebral , Núcleo Pulposo , Animais , Humanos , Bovinos , Hidrogéis/metabolismo , Sulfatos de Condroitina , Degeneração do Disco Intervertebral/tratamento farmacológico , Colágeno Tipo IIRESUMO
Visible light responsive micellar drug delivery formulations are of notable interest for the treatment of ocular diseases, as their successful development would enable controlled drug release at the back of the eye, improving efficacy and reducing side-effects when compared to existing approaches. In this work, an aliphatic polycarbonate-based visible light responsive micelle formulation based on mPEG-b-poly(5-hydroxy-trimethylene carbonate) (PHTMC) was prepared wherein the pendant hydroxyl groups of the PHTMC repeating units were protected by blue light-labile [7-(diethylamino)coumarin-4-yl]methyl (DEACM). The photo-labile DEACM provided a photo-triggered release profile, as, upon the removal of these protecting groups by photo-irradiation, the micelles underwent structural disruption, leading to the release of the payload. The removal of DEACM also deprotected the pendant hydroxyl groups of PHTMC, leading to PHTMC backbone degradation via intramolecular cyclization.
Assuntos
Corticosteroides , Micelas , Luz , Olho , Liberação Controlada de FármacosRESUMO
Current engineered synthetic scaffolds fail to functionally repair and regenerate ruptured native tendon tissues, partly because they cannot satisfy both the unique biological and biomechanical properties of these tissues. Ideal scaffolds for tendon repair and regeneration need to provide porous topographic structures and biological cues necessary for the efficient infiltration and tenogenic differentiation of embedded stem cells. To obtain crimped and porous scaffolds, highly aligned poly(l-lactide) fibers were prepared by electrospinning followed by postprocessing. Through a mild and controlled hydrogen gas foaming technique, we successfully transformed the crimped fibrous mats into three-dimensional porous scaffolds without sacrificing the crimped microstructure. Porcine derived decellularized tendon matrix was then grafted onto this porous scaffold through fiber surface modification and carbodiimide chemistry. These biofunctionalized, crimped, and porous scaffolds supported the proliferation, migration, and tenogenic induction of tendon derived stem/progenitor cells, while enabling adhesion to native tendons. Together, our data suggest that these biofunctionalized scaffolds can be exploited as promising engineered scaffolds for the treatment of acute tendon rupture.
Assuntos
Materiais Biocompatíveis , Teste de Materiais , Regeneração , Tendões , Alicerces Teciduais , Alicerces Teciduais/química , Tendões/citologia , Animais , Suínos , Porosidade , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Engenharia Tecidual , Proliferação de Células/efeitos dos fármacos , Tamanho da Partícula , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/farmacologia , Poliésteres/químicaRESUMO
The potential of a viscous liquid injectable delivery system composed of poly(5-ethylene ketal ε-caprolactone-co-D,L-lactide) (PEKCDLLA) to release bioactive vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) using an osmotic pressure release mechanism for the purpose of treating critical limb ischemia was investigated. VEGF and HGF were lyophilized separately with trehalose and bovine serum albumin (BSA) and incorporated into the polymer by simple mixing. VEGF and HGF were released by convective flow through superhydrated regions formed within the polymer as a result of the osmotic activity generated upon dissolution of the particles, along with the contributions of polymer degradation at later time points. A sustained release of highly bioactive VEGF and HGF for over 40 days with minimal burst was achieved under conditions of multidirectional delivery. The solubility of the growth factors in the concentrated trehalose solution formed upon dissolution of the particle within the polymer was determined to be a key parameter governing the rate and extent of growth factor release. This formulation approach, of using a low viscosity polymer delivery vehicle, is potentially useful for localized delivery of acid and temperature sensitive proteins, such as VEGF and HGF. This system may also serve as a platform for controlled and predictable delivery patterns for other therapeutic proteins in other clinical settings.
Assuntos
Fator de Crescimento de Hepatócito/farmacologia , Poliésteres/química , Proteínas Recombinantes/farmacologia , Fator A de Crescimento do Endotélio Vascular/química , Fator A de Crescimento do Endotélio Vascular/farmacologia , Animais , Bovinos , Química Farmacêutica/métodos , Sistemas de Liberação de Medicamentos/métodos , Extremidades , Liofilização/métodos , Fator de Crescimento de Hepatócito/química , Humanos , Concentração de Íons de Hidrogênio , Isquemia/tratamento farmacológico , Osmose/efeitos dos fármacos , Polímeros/química , Proteínas Recombinantes/química , Soroalbumina Bovina/química , Solubilidade , ViscosidadeRESUMO
High modulus, two-phase, bicontinuous scaffolds were prepared by photocross-linking an aqueous suspension of chondrocytes and N-methacrylate glycol chitosan with a hydrolyzable, hydrophobic, acrylated star-copolymer. Two acrylated star-copolymers were examined: poly(ε-caprolactone-co-d,l-lactide) (5446DLLACL) and poly(ε-caprolactone-co-trimethylene carbonate) (7030TMCCL). The scaffolds were assessed for injectability, two-phase interconnectivity, fatigue resistance, and long-term static culture behavior. The 7030TMCCL scaffolds demonstrated decreased moduli of 17% after 1 × 10(6) cycles at 30% strain and 5% after 56 days in culture, compared to the 5446DLLACL scaffolds, which exhibited decreases of 58 and 68%, respectively. The 7030TMCCL scaffolds accumulated more extracellular matrix after 56 days of culture (GAG: 20.1 ± 1, collagen: 35.5 ± 1.8 µg) compared to 5446DLLACL scaffolds (GAG: 13.2 ± 0.6, collagen: 6.2 ± 3.4 µg). Overall, the 7030TMCCL-based scaffolds were shown to be better suited for use as a load bearing soft tissue scaffold.
Assuntos
Lactonas/química , Poliésteres/química , Alicerces Teciduais/química , Animais , Materiais Biocompatíveis/química , Cartilagem/citologia , Cartilagem/fisiologia , Bovinos , Células Cultivadas , Condrócitos/fisiologia , Módulo de Elasticidade , Glicosaminoglicanos/metabolismo , Injeções , Teste de Materiais , Regeneração , Suporte de CargaRESUMO
Degenerative intervertebral disc disease is a common source of chronic pain and reduced quality of life in people over the age of 40. While degeneration occurs throughout the disc, it most often initiates in the nucleus pulposus (NP). Minimally invasive delivery of NP cells within hydrogels that can restore and maintain the disc height while regenerating the damaged NP tissue is a promising treatment strategy for this condition. Towards this goal, a biohybrid ABA dimethacrylate triblock copolymer was synthesized, possessing a lower critical solution temperature below 37 °C and which contained as its central block an MMP-degradable peptide flanked by poly(trimethylene carbonate) blocks bearing pendant oligoethylene glycol groups. This triblock prepolymer was used to form macroporous NP cell-laden hydrogels via redox initiated (ammonium persulfate/sodium bisulfite) crosslinking, with or without the inclusion of thiolated chondroitin sulfate. The resulting macroporous hydrogels had water and mechanical properties similar to those of human NP tissue and were mechanically resilient. The hydrogels supported NP cell attachment and growth over 28 days in hypoxic culture. In hydrogels prepared with the triblock copolymer but without the chondroitin sulfate the NP cells were distributed homogeneously throughout in clusters and deposited collagen type II and sulfated glycosaminoglycans but not collagen type I. This hydrogel formulation warrants further investigation as a cell delivery vehicle to regenerate degenerated NP tissue. STATEMENT OF SIGNIFICANCE: The intervertebral disc between the vertebral bones of the spine consists of three regions: a gel-like central nucleus pulposus (NP) within the annulus fibrosis, and bony endplates. Degeneration of the intervertebral disc is a source of chronic pain in the elderly and most commonly initiates in the NP. Replacement of degenerated NP tissue with a NP cell-laden hydrogel is a promising treatment strategy. Herein we demonstrate that a crosslinkable polymer with a lower critical solution temperature below 37 °C can be used to form macroporous hydrogels for this purpose. The hydrogels are capable of supporting NP cells, which deposit collagen II and sulfated glycosaminoglycans, while also possessing mechanical properties matching those of human NP tissue.
RESUMO
Specialized pro-resolving mediators (SPMs) are being considered for the treatment of chronic inflammatory diseases. However, these polyunsaturated fatty acids are prone to oxidation and as a result have a short biological half-life. It was reasoned that a micelle formulation would provide sustained delivery of SPMs while providing protection from oxidation. Thus, micelle formulations were prepared with poly(ethylene glycol) (PEG) as the hydrophilic block and poly(trimethylene carbonate) (PT) containing unsaturated pendant groups, specifically benzyloxy (BT) and sorbate (ST) groups, as the hydrophobic block. The potential of these micelles was assessed using linoleic acid as a model SPM. Loading into a micelle core reduced the extent of oxidation of the model SPM and a sustained release of non-oxidized model drug was achieved for up to 20 days in vitro from the PEG-P(T-BT) micelles. These micelles were also non-cytotoxic over a wide concentration range, demonstrating the potential of this formulation for effective SPM release in vivo.
Assuntos
Portadores de Fármacos , Micelas , Portadores de Fármacos/química , Polímeros/química , Polietilenoglicóis/química , Interações Hidrofóbicas e HidrofílicasRESUMO
Low molecular weight poly(5-ethylene ketal ε-caprolactone-co-D,L-lactide) (PEKCDLLA) is being considered as a viscous liquid, injectable depot for localized drug delivery. This polymer degrades in vitro via surface erosion, which is potentially advantageous for the proposed application. However, the in vivo degradation rate and mechanism, and tissue response, to polymers based on 5-ethylene ketal ε-caprolactone have not yet been reported. The purpose of this study was to measure the in vivo weight loss and change in polymer properties and assess the tissue response to PEKCDLLA after subcutaneous injection in rats. The tissue response was assessed histologically using Masson's trichrome staining and immunohistochemically by staining for CD68 positive cells. The polymer lost weight with time in a nearly linear fashion but did not exhibit significant changes in number average molecular weight, polydispersity index, and glass transition temperature or monomer ratio, consistent with a surface erosion process. The tissue response to the polymer was moderate and comparable to that reported in the literature for other degradable polymers used in clinical applications. These findings indicate that PEKCDLLA is a promising candidate for injectable drug delivery.
Assuntos
Implantes Absorvíveis , Portadores de Fármacos/farmacocinética , Poliésteres/farmacocinética , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Portadores de Fármacos/toxicidade , Fibroblastos/metabolismo , Células Gigantes de Corpo Estranho/metabolismo , Células Gigantes de Corpo Estranho/patologia , Inflamação/induzido quimicamente , Inflamação/patologia , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Poliésteres/toxicidade , Ratos , Ratos Wistar , Tela Subcutânea/efeitos dos fármacos , Tela Subcutânea/patologia , Temperatura de TransiçãoRESUMO
The coencapsulation of growth factor-loaded microspheres with adipose-derived stem cells (ASCs) within a hydrogel matrix was studied as a potential means to enhance ASC chondrogenesis in the development of a cell-based therapeutic strategy for the regeneration of partial thickness chondral defects. A photopolymerizable N-methacrylate glycol chitosan (MGC) was employed to form an in situ gel used to encapsulate microspheres loaded with bone morphogenetic protein 6 (BMP-6) and transforming growth factor-ß3 (TGF-ß3) with human ASCs. ASC viability and retention were enhanced when the Young's modulus of the MGC ranged between 225 and 380 kPa. Grafting an RGD-containing peptide onto the MGC backbone (RGD-MGC) improved ASC viability within the gels, remaining at greater than 90% over 14 days in culture. The effects of BMP-6 and TGF-ß3 released from the polymer microspheres on ASC chondrogenesis were assessed, and the level of differentiation was compared to ASCs in control gels containing nongrowth factor-loaded microspheres cultured with and without the growth factors supplied in the medium. There was enhanced expression of chondrogenic markers at earlier time points when the ASCs were induced with the sustained and local release of BMP-6 and TGF-ß3 from the microspheres. More specifically, the normalized glycosaminoglycan and collagen type II protein expression levels were significantly higher than in the controls. In addition, the ratio of collagen type II to type I was significantly higher in the microsphere delivery group and increased over time. End-point RT-PCR analysis supported that there was a more rapid induction and enhancement of ASC chondrogenesis in the controlled release group. Interestingly, in all of the assays, there was evidence of chondrogenic differentiation when the ASCs were cultured in the gels in the absence of growth factor stimulation. Overall, the co-delivery of growth-factor-loaded microspheres and ASCs in RGD-modified MGC gels successfully induced ASC chondrogenesis and is a promising strategy for cartilage repair.
Assuntos
Células-Tronco Adultas/fisiologia , Cartilagem Articular/fisiologia , Quitosana/análogos & derivados , Quitosana/síntese química , Hidrogéis/síntese química , Metacrilatos/síntese química , Engenharia Tecidual , Tecido Adiposo Branco/citologia , Células-Tronco Adultas/metabolismo , Células-Tronco Adultas/transplante , Sequência de Aminoácidos , Cápsulas/química , Cápsulas/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Quitosana/toxicidade , Condrogênese , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Módulo de Elasticidade , Feminino , Expressão Gênica , Humanos , Hidrogéis/toxicidade , Metacrilatos/toxicidade , Microesferas , Oligopeptídeos/química , Polimerização/efeitos da radiação , RegeneraçãoRESUMO
Liquid, injectable hydrophobic polymers are potentially useful as depot systems for localized drug delivery. Low molecular weight polymers of 5-ethylene ketal ε-caprolactone and copolymers of this monomer with D,L-lactide were prepared and their properties assessed with respect to their suitability for this purpose. The polymers were amorphous and of low viscosity, and the viscosity was adjustable by choice of initiator and/or by copolymerizing with D,L-lactide. Lower viscosity polymers were attained by using 350 Da methoxy poly(ethylene glycol) as an initiator in comparison to octan-1-ol, while copolymerization with D,L-lactide increased viscosity. The initiator used had no significant effect on the rate of mass loss in vitro, and copolymers with D,L-lactide (DLLA) degraded faster than 5-ethylene ketal ε-caprolactone (EKC) homopolymers. For the EKC-based polymers, a nearly constant degradation rate was observed. This finding was attributed to the hydrolytic susceptibility of the EKC-EKC ester linkage, which was comparable to that of DLLA-DLLA, coupled with a higher molecular weight of the water-soluble degradation product and the low initial molecular weight of the EKC-based polymers. Cytotoxicity of the hydrolyzed EKC monomer to 3T3 fibroblast cells was comparable to that of ε-caprolactone, suggesting that polymers prepared from EKC may be well tolerated upon in vivo implantation.
Assuntos
Materiais Biocompatíveis/síntese química , Caproatos/química , Preparações de Ação Retardada/síntese química , Injeções/métodos , Lactonas/química , Poliésteres/síntese química , Células 3T3 , Aldeídos/química , Animais , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Preparações de Ação Retardada/farmacologia , Estabilidade de Medicamentos , Ésteres/química , Teste de Materiais , Camundongos , Peso Molecular , Poliésteres/farmacologia , Polietilenoglicóis/química , Propriedades de Superfície , Viscosidade , ÁguaRESUMO
A recently developed viscous liquid aliphatic polycarbonate, poly(trimethylene carbonate-co-5-hydroxy trimethylene carbonate), has advantageous properties for the delivery of acid-sensitive drugs such as proteins and peptides. This copolymer degrades in vitro via an alkaline-catalyzed intramolecular cyclization reaction yielding oligo (trimethylene carbonate), glycerol, and carbon dioxide, but its in vivo degradation mechanisms are presently unknown. The in vivo degradation mechanism and tissue response to this copolymer were investigated following subcutaneous implantation in Wistar rats. The molecular weight and composition of the copolymer varied in the same manner following subcutaneous implantation as observed in vitro. These findings suggest that the copolymer also degraded in vivo principally via intramolecular cyclization. The tissue response in terms of the inflammatory zone cell density, fibrous capsule thickness, and macrophage response was intermediate to that of two clinically used biodegradable sutures, Vicryl and Monocryl, indicating that the copolymer can be considered biotolerable. Collectively, the data show that further development of this copolymer as a drug delivery material is warranted.
Assuntos
Materiais Biocompatíveis/metabolismo , Cimento de Policarboxilato/metabolismo , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Catálise , Ciclização , Portadores de Fármacos/química , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Cimento de Policarboxilato/química , Cimento de Policarboxilato/farmacologia , Próteses e Implantes , Ratos , Ratos WistarRESUMO
Semicrystalline poly(l-lactide-co-ε-caprolactone) (P(LLA-CL)) was used to produce electrospun fibers with diameters on the subcellular scale. P(LLA-CL) was chosen because it is biocompatible and its chemical and physical properties are easily tunable. The use of a rotating wire mandrel as a collection device in the electrospinning process, along with high collection speeds, was used to align electrospun fibers. Upon removal of the fibers from the mandrel, the fibers shrunk in length, producing a crimp pattern characteristic of collagen fibrils in soft connective tissues. The crimping effect was determined to be a result of the residual stresses resident in the fibers due to the fiber alignment process and the difference between the operating temperature (T(op)) and the glass-transition temperature (T(g)) of the polymer. The electrospun fibers could be induced to crimp by adjusting the operating temperature to be greater than that of the polymer glass-transition temperature. Moreover, the crimped fibers exhibited a toe region in their stress-strain profile that is characteristic of collagen present in tendons and ligaments. The crimp pattern was retained during in vitro degradation over 4 weeks. Primary bovine fibroblasts seeded onto these crimped fibers attached, proliferated, and deposited extracellular matrix (ECM) molecules on the surface of the fiber mats. These self-crimping fibers hold great promise for use in tissue engineering scaffolds for connective tissues that require fibers similar in structure to that of crimped collagen fibrils.
Assuntos
Poliésteres/uso terapêutico , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Materiais Biocompatíveis , Bovinos , Técnicas de Cultura de Células , Tecido Conjuntivo/crescimento & desenvolvimento , Fibroblastos/citologiaRESUMO
Polymeric biohybrid networks have significant potential as supportive materials for soft connective tissue regeneration. Their success in this regard is determined by their initial mechanical properties, which are dependent on their water content, as well as the rate at which these properties change with time due to cell mediated degradation. In this study the in vivo degradation and tissue response following implantation of matrix metalloproteinase (MMP)-degradable poly(trimethylene carbonate) (PTMC)-based biohybrid networks were assessed in a Wistar rat model. The networks examined varied in equilibrium water content from circa 20% to 70% w/w. The networks degraded through MMP secretion by inflammatory cells at the tissue-material interface, generating a mass loss profile consistent with surface erosion but modulus and sol content changes consistent with a bulk erosion process. This degradation profile was explained in terms of a population gradient in MMP concentration from the surface to the bulk of the networks due to diffusion restrictions. A histological analysis of the tissue surrounding the implants confirmed a moderate tissue response comparable to that observed towards a VicrylTM suture, suggesting that these new materials can be considered biocompatible.
Assuntos
Dioxanos/química , Enzimas/química , Metaloproteinases da Matriz/química , Polímeros/química , Água/química , Animais , Materiais Biocompatíveis/química , Difusão , Inflamação , Masculino , Teste de Materiais , Peptídeos/química , Ratos , Ratos Wistar , Regeneração , Propriedades de Superfície , SuturasRESUMO
Viscous liquid degradable polymers have advantages as drug depots for sustained protein delivery. We have created a new aliphatic polycarbonate for this purpose, poly(trimethylene carbonate-co-5-hydroxy trimethylene carbonate), which upon degradation retains a near neutral micro-environmental pH. As such, this copolymer is highly suited to the delivery of acid sensitive proteins. We show that the mechanism of protein release from this liquid copolymer is consistent with the formation of super-hydrated regions as a result of the osmotic activity of the solution formed upon distributed protein particle dissolution. Protein release can be manipulated by controlling polymer hydrophobicity which can be adjusted by molecular weight and choice of initiator. Moreover, protein release is highly dependent on protein solubility which impacts the osmotic activity of the solution formed upon dissolution of the protein particles while protein molecular size and isoelectric point are not as influential. As demonstrated by the release of highly bioactive vascular endothelial growth factor, formulations of this copolymer are suitable for prolonged delivery of protein therapeutics.
Assuntos
Polímeros , Fator A de Crescimento do Endotélio Vascular , Sistemas de Liberação de Medicamentos , Cimento de Policarboxilato , ViscosidadeRESUMO
Biohybrid networks have the potential to have stiffnesses equivalent to that of native soft connective tissues as well as cell-mediated degradation behavior. Most strategies to generate such materials to date have utilized crosslinking of two separate and orthogonally functionalized polymers. Herein we describe a triblock prepolymer consisting of a central enzyme degradable peptide block flanked by two synthetic, hydrolysis resistant poly(trimethylene carbonate) blocks (PTMC) or poly(ethylene glycol)-PTMC blocks terminated in methacrylate groups. To form these prepolymers heterobifunctional PTMC and PEG-PTMC were prepared, possessing a vinyl sulfone terminus and a methacrylate terminus. These polymers were conjugated to a di-cysteine containing peptide through a Michael-type addition to form cross-linkable prepolymers. These prepolymers were then photo-cured to form enzyme degradable networks. The compressive moduli of the resulting water swollen networks was within the range of many soft connective tissues and was inversely proportional to the water solubility of the prepolymers. The prepolymer water solubility in turn could be tuned by adjusting PTMC molecular weight or by the addition of a PEG block. In vitro degradation only occurred in the presence of matrix metalloproteinases, and was fastest for networks prepared with prepolymers of higher water solubility.