Polyurethane scaffold formation via a combination of salt leaching and thermally induced phase separation.

Porous scaffolds have been made from two polyurethanes based on thermally induced phase separation of polymer dissolved in a DMSO/water mixture in combination with salt leaching. It is possible to obtain very porous foams with a very high interconnectivity. A major advantage of this method is that variables like porosity, pore size, and interconnectivity can be independently adjusted with the absence of toxic materials in the production process. The obtained compression moduli were between 200 kPa and 1 MPa with a variation in porosity between 76 and 84%. Currently the biological and medical aspects are under evaluation.

Meniscal replacement in dogs. Tissue regeneration in two different materials with similar properties.

In earlier studies, meniscal replacement with a porous polymer implant led to regeneration of neo-meniscal tissue. To evaluate the influence of the chemical properties on the tissue regeneration in the implant, in the present study, the meniscus in the dog's knee was replaced with either an aromatic 4,4-diphenylmethanediisocyanate based polyesterurethane implant (Estane) (n = 6) or with an aliphatic 1,4-butanediisocyanate based polyesterurethane implant (PCLPU) (n = 6). After 6 months, the knee joints were resected and the tissue behavior in the two different prostheses was evaluated microscopically. In both prostheses, a meniscus-like distribution of the tissue phenotype was found with collagen type I in the peripheral fibrous zones and collagen type II in the central, more cartilaginous zones. The compression-stress behavior of the implant-tissue construct remained in between the stiffness of the polymer material and that of the native meniscus. The PCLPU implant seemed to provoke less synovial tissue reaction. After meniscectomy solely, in 5 out of 6 cases, a meniscus-like regenerate was formed. Furthermore, the articular cartilage degeneration after placing a PCLPU implant did also not exceed the degeneration after the Estane implant or after meniscectomy. The differences between these two implants did not seem to influence the tissue regeneration in the implant. However, PCLPU seemed to evoke less tissue reaction and, therefore, is thought to be less or even nontoxic as compared with the Estane implant. Therefore, for studies in the future, the authors prefer the PCLPU prostheses for replacement of the meniscus.

Design, synthesis and properties of a degradable polyurethane scaffold for meniscus regeneration.

Longitudinal lesions in menisci are among the most frequent orthopedic problems of the knee. Repair by simple techniques is only limited to the vascular part of the meniscus. For repair of the avascular part of the meniscus a scaffold, which will assist the body in the formation of new meniscus cell tissue, might be applicable. In this study a biomedical segmented polyurethane with poly(epsilon-caprolactone) as soft segment and 1,4-butanediisocyanate and 1,4-butanediol as uniform hard segments has been synthesised. The material has a micro phase separated morphology and excellent mechanical properties. A porous scaffold was prepared via a combination of liquid-liquid phase separation and salt leaching. The foams prepared combined a very high interconnectivity and porosity with the desired compression modulus. After six months of implantation in the knees of beagles full ingrowth with cells was obtained and it was found that meniscus like tissue had been formed in the scaffold. Moreover, compression behaviour appeared to be comparable to native meniscus tissue.

A porous polymer scaffold for meniscal lesion repair--a study in dogs.

Meniscal lesions often occur in the avascular area of the meniscus with little chance of spontaneous repair. An access channel in the meniscal tissue can function as an entrance for ingrowing repair tissue from the vascular periphery of the meniscus to the lesion in the avascular zone which again induced healing of the lesion. Implantation of a porous polymer in a full-thickness access channel induced healing. However, a better integration between meniscal tissue and the implant might be achieved with the combination of the newly developed porous polymers and a modified surgical technique. This might improve meniscal lesion healing and the repair of the access channel with neo-meniscal tissue. Longitudinal lesions were created in the avascular part of 24 canine lateral menisci and a partial-thickness access channel was formed to connect the lesion with the meniscal periphery. In 12 menisci, the access channel was left empty (control group), while in the remaining 12 menisci the polymer implant was sutured into the access channel. Repair of the longitudinal lesions was achieved with and without polymer implantation in the partial-thickness access channel. Polymer implants induced fibrous ingrowth with cartilaginous areas, which resembled neo-meniscal tissue. Implantation did not prevent articular cartilage degeneration.

Presence and mechanism of knee articular cartilage degeneration after meniscal reconstruction in dogs.

OBJECTIVE: Partial meniscectomy is the golden standard for treating a bucket-handle tear in the meniscus of the knee, but it inevitably leads to articular cartilage degeneration. Surgical creation of an access channel between the lesion and the vascularized synovial lining is intended to induce ingrowth of repair tissue and thus avoid degeneration of articular cartilage. DESIGN: The presence and mechanism of cartilage degeneration were evaluated in 24 canine menisci after a longitudinal lesion and access channel had been created in the avascular part of the meniscus. In 12 menisci the channel was implanted with a porous polymer scaffold, while the remaining 12 were left empty. Evaluation was performed using routine histology and antibodies directed against denatured type II collagen (Col2-3/4M). RESULTS: Articular degeneration was apparent in the polymer implant group and the empty channel group. This consisted of fibrillation, loss of chondrocytes and decreased proteoglycan content. Areas of fibrillated cartilage always showed positive labeling with the collagen degradation antibody Col2-3/4M. Collagen degradation was also visible in non-fibrillated areas. The upper zone of the cartilage showed swelling especially in the implant group, with empty cell lacunae and moderate levels of Col2-3/4M antibody labeling. DISCUSSION: This reconstruction technique cannot be considered superior to partial meniscectomy. We propose that degradation of the collagen type II network is a result of cartilage fibrillation and vice versa.

Cytotoxicity of poly(96L/4D-lactide): the influence of degradation and sterilization.

The cytotoxicity of poly(96L/4D-lactide) (PLA96), and of its accumulated degradation products, was investigated following different sterilization methods and pre-determined heat-accelerated degradation intervals. PLA96 samples sterilized by either steam, ethylene oxide, or gamma irradiation were left untreated (S0 samples), or were degraded for 30 h or 60 h (S30 and S60 samples) at 90 degrees C in water. Extracts of the samples and of the remaining degradation fluids (F30 and F60) were prepared. The toxicity of both unfiltered and filtered extracts was analyzed in a cell growth inhibition (CGI) assay and a lactate dehydrogenase (LDH) leakage assay. Physical analysis of the extracted samples and of the degradation fluids also was performed. The S0 extracts demonstrated no significant CGI. The CGI of the S30 extracts ranged from 37 to 78%, whereas the CGI of the S60 extracts ranged from 6 to 33%. The CGI of the F30 extracts ranged from 19 to 38% and the CGI of the F60 extracts was 98 to 123%. The LDH leakage assay only showed a high response to the unfiltered F60 extracts. Neither sterilization nor filtration appeared to influence the cytotoxicity of the extracts. Particle accumulation, however, might affect cell membrane permeability resulting in LDH leakage. The results of this study suggest that the cytotoxicity of PLA96 is related to the pH and possibly the osmolarity of the tested extracts. The pH and osmolarity, in turn, may depend on variations in the amounts of solubilized lactic acid and oligomers. These variations appear to result from degradation stage-dependent differences in crystallinity, molecular weight and molecular weight distribution of the PLA96 samples.

Solvent-free fabrication of micro-porous polyurethane amide and polyurethane-urea scaffolds for repair and replacement of the knee-joint meniscus.

New porous polyurethane urea and polyurethane amide scaffolds for meniscal reconstruction have been developed in a solvent-free process. As soft segments, copolymers of 50/50 L-lactide/epsilon-caprolactone have been used. After terminating the soft segment with diisocyanates, chain extension was performed with adipic acid and water. Reaction between the isocyanate groups and adipic acid or water provides carbon dioxide and results in a porous polymer. Extra hydroxyl-terminated prepolymer was added in order to regulate the amount of carbon dioxide formed in the foaming reaction. Furthermore, salt crystals ranging in size from 150 to 355 microm were added in order to induce macroporosity. The pore size was regulated by addition of surfactant and by the use of ultrasonic waves. The resulting porous polymer scaffolds exhibit good mechanical properties like a high-compression modulus of 150 kPa. Chain extension with adipic acid results in better mechanical properties due to better defined hard segments. This results from the lower nucleophilicity of carboxylic acids compared to water and alcohols. By adjusting the reaction conditions, materials in which macropores are interconnected by micropores can be obtained. On degradation only non-toxic products will be released; importantly, the materials were obtained by a simple, reproducible and solvent-free procedure.

The effect of chromic acid treatment on the mechanical and tribological properties of aramid fibre reinforced ultra-high molecular weight polyethylene composite.

Surface oxidation of ultra-high molecular weight polyethylene (UHMWPE) powder has an influence on the mixing procedure of chopped fibres and UHMWPE powder. Due to this oxidation hydrogen bonds can be formed between the fibres and powder particles, leading to a more homogeneous fibre-powder mixture. This treatment improves the fibre-matrix interface and thus the physical properties of the composite. Chromic acid treatment also has an influence on the mechanical and tribological properties of the aramid-UHMWPE composite. Although only a relatively small improvement is observed in the modulus, yield stress and stress at break, of 33, 17 and 9%, respectively, a substantial enhancement in wear resistance of 117% is observed.

A new biomedical polyurethane with a high modulus based on 1,4-butanediisocyanate and epsilon-caprolactone.

A new approach to the synthesis of biomedical polyurethanes based on epsilon-caprolactone and 1,4-butanediisocyanate with a high modulus, has been developed. By chain extending an epsilon-caprolactone prepolymer with a long uniform-size diisocyanate block, a segmented polyurethane with uniform-size hard segments was obtained. It shows excellent mechanical properties; an extremely high modulus of 105 MPa and a tensile strength of 35 MPa. The polymer is soluble at high concentrations in various volatile solvents such as chloroform and 1,4-dioxane. By a combination of salt-leaching and freeze-drying, porous materials have been obtained in which macropores ranging in size from 150-300 microm are highly interconnected by micropores. The material shows a sufficiently high compression modulus of 200 kPa and appears to be suitable for biomedical applications such as meniscal prostheses.

Long-term evaluation of degradation and foreign-body reaction of subcutaneously implanted poly(DL-lactide-epsilon-caprolactone).

The aim of this study was to evaluate the degradation and foreign-body reaction of poly(DL-lactide-epsilon-caprolactone) (PLA85CL50) bars. This specific biomaterial is used for the construction of nerve guides, which can be used in the reconstruction of short nerve gaps. Subcutaneously implanted PLA85CL50 bars were harvested after implantation periods ranging from 3 to 12 months and evaluated for the rate of degradation and the degree of foreign-body reaction. It was observed that this copolymer degraded completely within 12 months and that no lactide or epsilon-caprolactone crystals were formed. Furthermore, we conclude that the foreign-body reaction of PLA85CL50 is very mild. These properties make the amorphous copolymer of DL-lactide and epsilon-caprolactone (50:50) suitable for the construction of nerve guides.

Evaluation of functional nerve recovery after reconstruction with a new biodegradable poly (DL-lactide-epsilon-caprolactone) nerve guide.

The aim of this study was to evaluate functional nerve recovery following reconstruction of a 1 cm gap in the sciatic nerve of a rat, using a new biodegradable p (DLLA-epsilon-CL) nerve guide. To evaluate both motor and sensory nerve recovery, walking track analysis and electrostimulation tests were carried out after implantation periods, ranging from 3 to 15 weeks post-operatively. The first signs of functional nerve recovery were observed after 3 weeks. After 15 weeks, 70% of the motor- and 90% of the sensory nerve function was re-established. Return of nerve function was better, in comparison with results from other studies. This study demonstrated successful functional nerve recovery after the reconstruction of a 1 cm nerve gap with a biodegradable p(DLLA-epsilon-CL) nerve guide.

Meniscal tissue regeneration in porous 50/50 copoly(L-lactide/epsilon-caprolactone) implants.

Porous materials of a high-molecular-weight 50/50 copolymer of L-lactide and epsilon-caprolactone with different compression moduli were used for meniscal repair. In contrast to the previously used 4,4'-diphenylmethane and 1,4-trans-cyclohexane diisocyanates containing polyurethanes, degradation products of the copolymer are non-toxic. Two series of porous materials with compression moduli of 40 and 100 kPa respectively were implanted in the knees of dogs using a new, less traumatizing suturing technique. A porous aliphatic polyurethane series with compression modulus of 150 kPa was implanted for comparison. Adhesion of the implant to meniscal tissue was found to be essential for healing of the longitudinal lesion. Copolymer implants showed better adhesion, probably due to the higher degradation rate of the copolymer. Fibrocartilage formation was found to be affected by the compression modulus of the implant. Implants with a modulus of 40 kPa did not show ingrowth of fibrocartilage, whereas implants with compression moduli of 100 and 150 kPa yielded 50-70 and 80-100% fibrocartilage respectively. During degradation the copolymer phase separated into a crystalline phase containing mainly L-lactide and an amorphous phase containing mainly epsilon-caprolactone. The copolymer degraded through bulk degradation.

A novel method for fabrication of biodegradable scaffolds with high compression moduli.

It has been previously shown that, when used for meniscal reconstruction, porous copoly(L-lactide/epsilon-caprolactone) implants enhanced healing of meniscal lesions owing to their excellent adhesive properties. However, it appeared that the materials had an insufficient compression modulus to accomplish 100% fibrocartilage formation. In addition, to be used for meniscal prosthesis, the compression modulus of the porous materials should be larger than 150 kPa in order to protect the articular cartilage. A technique was developed to prepare stiff porous materials of a high molecular weight 50/50 copoly(L-lactide/epsilon-caprolactone) suitable for fibrocartilage regeneration in meniscal implants and meniscal prosthesis. Porous microspheres (50-250 microm) were agglutinated in the presence of NaCl crystals (250-300 microm). The microspheres were mixed with solid solvent in order to obtain a homogeneous distribution of solvent over the spheres. By changing the amount of solvent and crystals, the density and the compression modulus could be varied over a range of 0.07 g ml(-1) to 0.5 g dl(-1) and 40-1100 kPa, respectively.

Meniscal repair by fibrocartilage in the dog: characterization of the repair tissue and the role of vascularity.

Lesions in the avascular part of 20 canine menisci were repaired by implantation of a porous polyurethane. Seven menisci were not repaired and served as controls. The repair tissue was characterized by biochemical and immunological analysis. The role of vascularity in healing was studied by perfusion of menisci with Indian ink. Histologically, repair tissue inside the implants initially consisted of fibrous tissue containing type I collagen. After 2 months, fibrocartilaginous tissue developed inside the implants, whereas control defects only showed repair with fibrous tissue. Both type I and type II collagen, the two major collagen types of normal meniscal fibrocartilage, could be detected in this newly formed fibrocartilage. The implant guided vascular tissue from the periphery towards the lesion resulting in healing of the tear. After fibrocartilage had formed, vascularity decreased and was completely absent in mature fibrocartilage. Control defects remained filled with vascular connective tissue. Two-thirds of the longitudinal lesions were found to be healed partially or completely. It is concluded that implantation of a porous polymer does enhance vascularity sufficiently to result in healing of meniscal lesions extending into the avascular part. Healing takes place by repair tissue strongly resembling normal meniscal fibrocartilage.

Meniscal replacement using a porous polymer prosthesis: a preliminary study in the dog.

A porous polyurethane prosthesis was used to replace the lateral meniscus in the dog. After an initial ingrowth of fibrous tissue, the prostheses became filled with tissue strongly resembling normal meniscal fibrocartilage. Although less severe than seen after total meniscectomy, cartilage degeneration was frequent, possibly because tissue ingrowth in the prostheses occurred too slowly. Porous polymers can be useful for replacement of the meniscus, provided that chemical and physical properties are optimized.

Light-microscopic and electron-microscopic evaluation of short-term nerve regeneration using a biodegradable poly(DL-lactide-epsilon-caprolacton) nerve guide.

The aim of this study was to evaluate short-term peripheral nerve regeneration across a 10-mm. gap, using a biodegradable poly(DL-lactide-epsilon-caprolacton) nerve guide, with an internal diameter of 1.5 mm and a wall thickness of 0.30 mm. To do so, we evaluated regenerating nerves using light microscopy, transmission electron microscopy and morphometric analysis after implantation of 12-mm nerve guides in the sciatic nerve of the rat. Evaluation times ranged from 3-10 weeks. Three weeks after reconstruction, myelinated nerve fibers could be observed in the distal nerve stump. Ten weeks after reconstruction, the regenerating nerves already resembled normal nerves. In conclusion, we show that poly(DL-lactide-epsilon-caprolacton) nerve guides can be successfully applied in the reconstruction of severed nerves in the rat model. Furthermore, we have observed the fastest nerve regeneration described thus far, after reconstruction using a biodegradable nerve guide.

Use of porous polyurethanes for meniscal reconstruction and meniscal prostheses.

In the past, porous materials made of an aromatic polyurethane (PU) were successfully used to meniscal reconstruction in dogs. Since aromatic PUs yield very toxic fragments upon degradation, a linear PU was synthesized by curing a poly(epsilon-caprolactone) and 1,4-trans-cyclohexane diisocyanate based prepolymer with cyclohexanedimethanol. Porous materials of this polymer were also implanted for meniscal reconstruction. The results were comparable with the most successful implant series so far. Additionally, a porous meniscal prosthesis was developed to replace a total meniscus. Due to the very high shear stresses to which the prosthesis would be exposed, the stress hysteresis phenomenon linear PUs are known to exhibit could be of great consequence. Therefore an aliphatic PU network, synthesized by cross-linking poly(epsilon-caprolactone) and 1,4-trans-cyclohexane diisocyanate with glycerol, was used. Dislocation caused by tearing out of the sutures was found to be a problem because the tear resistance of the material was relatively low. In this study the tearing problem has been partly circumvented by using a complex suturing technique. Meniscal prostheses turned out to induce fibrocartilage upon implantation, and degeneration of articular cartilage was less severe than after meniscectomy.

Poly(DL-lactide-epsilon-caprolactone) nerve guides perform better than autologous nerve grafts.

The aim of this study was to compare the speed and quality of nerve regeneration after reconstruction using a biodegradable nerve guide or an autologous nerve graft. We evaluated nerve regeneration using light microscopy, transmission electron microscopy and morphometric analysis. Nerve regeneration across a short nerve gap, after reconstruction using a biodegradable nerve guide, is faster and qualitatively better, when compared with nerve reconstruction using an autologous nerve graft. Therefore, we conclude that in the case of a short nerve gap (1 cm), reconstruction should be carried out using a biodegradable nerve guide constructed of a copolymer of DL-lactide and epsilon-caprolactone.

High-impact poly(L/D-lactide) for fracture fixation: in vitro degradation and animal pilot study.

The impact strength of amorphous lactide copolymers can be significantly improved by blending with biodegradable rubbers. Rubber toughening of amorphous poly(85L/15D -lactide) with the copolymer poly (50/50-trimethylenecarbonate-co-epsilon-caprolactone) results in a high-impact polymer (PDLLA/P(TMC-CL)). In vitro, the PDLLA/P(TMC-CL) blend retained its tensile and impact strength for a long period of time. Up to 45 weeks, the amount of water absorbed by the blend remained very low and no significant mass loss was observed. To test the suitability for fracture fixation, in a dog study mandibular fractures were fixated with PDLLA/P(TMC-CL) bone plates and screws. Bone healing was uneventful without premature failure of the implants. Although long-term degradation studies have to be carried out, PDLLA/P(TMC-CL) seems to be promising for application in fracture fixation.

Biological performance of a degradable poly(lactic acid-epsilon-caprolactone) nerve guide: influence of tube dimensions.

One of the ways to reconstruct a nerve defect is to use a biodegradable nerve guide. The aim of this study was to establish a nerve guide constructed of an amorphous copolymer of lactic acid-caprolactone. A pilot study was set up to elucidate the effect of the tube dimensions on nerve regeneration. Four types of nerve guides, with internal diameters ranging from 1.12-1.23 mm and wall thicknesses ranging from 0.34-0.68, were tested for this purpose. We evaluated the biodegradation, foreign body reaction and nerve regeneration by light microscopy, after three different implantation times (1, 2, and 3 months). After 2 months, we observed that all types of nerve guides had changed from a transparent to an opaque and swollen state, and that they had lost their strength. The foreign body reaction was characterized by the presence of giant cells and fibroblasts surrounding the degrading nerve guide. From this pilot study, we conclude that nerve guide type 1, with an internal diameter of 1.23 mm and a wall thickness of 0.34 mm, can ensure nerve regeneration in the case of a 1-cm gap in the sciatic nerve of the rat. Nerve guides types 3 and 4, with relatively small lumens, show nerve compression due to a more pronounced swelling of the degrading tube.