Personalized Fiber-Reinforcement Networks for Meniscus Reconstruction.

The menisci are fibrocartilaginous tissues that are crucial to the load-sharing and stability of the knee, and when injured, these properties are compromised. Meniscus replacement scaffolds have utilized the circumferential alignment of fibers to recapitulate the microstructure of the native meniscus; however, specific consideration of size, shape, and morphology has been largely overlooked. The purpose of this study was to personalize the fiber-reinforcement network of a meniscus reconstruction scaffold. Human cadaveric menisci were measured for a host of tissue (length, width) and subtissue (regional widths, root locations) properties, which all showed considerable variability between donors. Next, the asymmetrical fiber network was optimized to minimize the error between the dimensions of measured menisci and predicted fiber networks, providing a 51.0% decrease (p = 0.0091) in root-mean-square (RMS) error. Finally, a separate set of human cadaveric knees was obtained, and donor-specific fiber-reinforced scaffolds were fabricated. Under cyclic loading for load-distribution analysis, in situ implantation of personalized scaffolds following total meniscectomy restored contact area (253.0 mm2 to 488.9 mm2, p = 0.0060) and decreased contact stress (1.96 MPa to 1.03 MPa, p = 0.0025) to near-native values (597.4 mm2 and 0.83 MPa). Clinical use of personalized meniscus devices that restore physiologic contact stress distributions may prevent the development of post-traumatic osteoarthritis following meniscal injury.

Achieving molecular orientation in thermally extruded 3D printed objects.

Three-dimensional (3D) printing is used to fabricate tissue scaffolds. Polymer chains in these objects are typically unoriented. The mechanical properties of these scaffolds can be significantly enhanced by proper alignment of polymer chains. However, post-processing routes to increase orientation can be limited by the geometry of the printed object. Here, we show that it is possible to orient polymer chains during printing by optimizing printing parameters to take advantage of the flow characteristics of the polymer. This is demonstrated by printing a polymeric scaffold for meniscus regeneration using poly(desaminotyrosyl-tyrosine dodecyl dodecanedioate), poly(DTD DD). Alignment of polymer chains was achieved by translating the printhead at sufficiently high speeds when the polymer was still in a semi-solid state as it cooled from the fluid state at the tip of the nozzle using a critical combination of nozzle diameter, extrusion pressure, and temperature. The degree of orientation as evaluated by x-ray diffraction and thermal shrinkage, was greater than that of drawn fibers. Significant orientation and defect-free printing was achieved even for scaffolds with complex geometries. The ability to orient polymers during 3D printing has the potential to combine the advantages of 3D printing with the superior mechanical performance of more conventional polymer processing methods, such as drawing.

Biomechanical characterization of a novel collagen-hyaluronan infused 3D-printed polymeric device for partial meniscus replacement.

The menisci transmit load by increasing the contact area and decreasing peak contact stresses on the articular surfaces. Meniscal lesions are among the most common orthopedic injuries, and resulting meniscectomies are associated with adverse polycaprolactone contact mechanics changes and, ultimately, an increased likelihood of osteoarthritis. Meniscus scaffolds were fabricated by 3D-printing a network of circumferential and radial filaments of resorbable polymer (poly(desaminotyrosyl-tyrosine dodecyl ester dodecanoate)) and infused with collagen-hyaluronan. The scaffold demonstrated an instantaneous compressive modulus (1.66 ± 0.44 MPa) comparable to native meniscus (1.52 ± 0.59 MPa). The scaffold aggregate modulus (1.33 ± 0.51 MPa) was within 2% of the native value (1.31 ± 0.36 MPa). In tension, the scaffold displayed a comparable stiffness to native tissue (127.6-97.1 N/mm) and an ultimate load of 33% of the native value. Suture pull-out load of scaffolds (83.1 ± 10.0 N) was within 10% of native values (91.5 ± 15.4 N). Contact stress analysis demonstrated the scaffold reduced peak contact stress by 60-67% and increased contact area by 38%, relative to partial meniscectomy. This is the first meniscal scaffold to match both the axial compressive properties and the circumferential tensile stiffness of the native meniscus. The improvement of joint contact mechanics, relative to partial meniscectomy alone, motivates further investigation using a large animal model. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B:2457-2465, 2019.

Partial Meniscus Replacement with a Collagen-Hyaluronan Infused Three-Dimensional Printed Polymeric Scaffold.

IMPACT STATEMENT: The only FDA-approved partial meniscus scaffold, the Collagen Meniscus Implant (CMI), is not approved for reimbursement by government and only reimbursable by certain private insurers. Scaffolds with improved mechanical properties and greater efficacy are needed. A previous study (Ghodbane, et al. DOI: 10.1002/jbm.b.34331) demonstrated the ability of our novel acellular, off-the shelf scaffold to restore knee biomechanics following partial meniscectomy, which could potentially decrease the risk of osteoarthritis following partial meniscectomy, providing the motivation for this study. This article presents a first-in-animal feasibility study.

Tissue-Engineered Total Meniscus Replacement With a Fiber-Reinforced Scaffold in a 2-Year Ovine Model.

BACKGROUND: Meniscus injuries and associated meniscectomies cause patients long-term pain and discomfort and can lead to joint deterioration. PURPOSE: To evaluate a collagen-hyaluronan sponge reinforced with synthetic resorbable polymer fiber for total meniscus reconstruction in a long-term ovine model. STUDY DESIGN: Controlled laboratory study. METHODS: Eleven skeletally mature sheep were implanted with the total meniscus scaffold. At 2 years, explants were evaluated biologically (radial/circumferential histology, immunofluorescence) and mechanically (compression, tension), and articular surfaces were examined for damage. RESULTS: The fiber-reinforced scaffold induced formation of functional neomeniscus tissue that was intact in 8 of 11 animals. The implant was remodeled into organized circumferentially aligned collagen bundles to resist meniscus hoop stresses. Moreover, type II collagen and proteoglycan deposition near the inner margin suggested a direct response to compressive stresses and confirmed fibrocartilage formation. Cartilage damage was observed, but end-stage (severe) joint deterioration associated with meniscectomy was avoided, even with limitations regarding the ovine surgical procedure and postoperative care. CONCLUSION: A fiber-reinforced total meniscus replacement device induces formation of functional neomeniscus tissue that has the potential to prevent catastrophic joint deterioration associated with meniscectomy. CLINICAL RELEVANCE: An off-the-shelf meniscus device that can be remodeled into functional tissue and thus prevent or delay the onset of osteoarthritis could address a widespread clinical need after meniscus injury.

Carbodiimide cross-linking counteracts the detrimental effects of gamma irradiation on the physical properties of collagen-hyaluronan sponges.

Collagen-based scaffolds are extensively used in biomaterials and tissue engineering applications. These scaffolds have shown great biocompatibility and versatility, but their relatively low mechanical properties may limit use in orthopaedic load-bearing applications. Moreover, terminal sterilization with gamma irradiation, as is commonly performed with commercial devices, presents concerns over structural integrity and enzymatic stability. Therefore, the goal of this study was to test the hypothesis that EDC/NHS cross-linking (10 mM/5 mM) can protect collagen-hyaluronan sponges from the damaging effects of gamma irradiation. Specifically, we evaluated compressive and tensile mechanical properties, enzymatic stability, porosity and pore size, and swelling ratio. Ultimate tensile strength and elastic modulus exhibited increases (168.5 and 245.8%, respectively) following irradiation, and exhibited over tenfold increases (1049.2 and 1270.6%, respectively) following cross-linking. Irradiation affected pore size (38.4% decrease), but cross-linking prior to irradiation resulted in only a 17.8% decrease. Cross-linking also showed an offsetting effect on the equilibrium modulus, enzymatic stability, and swelling ratio of sponges. These results suggest that carbodiimide cross-linking of collagen-hyaluronan sponges can mitigate the structural damage typically experienced during gamma irradiation, warranting their use in tissue engineering applications.

* The Ovine Model for Meniscus Tissue Engineering: Considerations of Anatomy, Function, Implantation, and Evaluation.

Meniscus injuries represent one of the most-common intra-articular knee injuries. The current treatment options include meniscectomy and allograft transplantation, both with poor long-term outcomes. Therefore, there is a need for regenerative techniques to restore meniscal function. To preclinically test scaffolds for meniscus replacement, large animal models need to be established and standardized. This review establishes the anatomical and compositional similarities between human and sheep menisci and provides guidance for implantation and evaluation of such devices. The ovine meniscus represents a scaled-down version of the human meniscus, with only slight structural differences that can be addressed during device fabrication. Implantation protocols in sheep remain a challenge, as the meniscus cannot be visualized with the arthroscopic-assisted procedures commonly performed in human patients. Thus, we recommend the appropriate implantation protocols for meniscus visualization, ligamentous restoration, and surgical fixation of both total and partial meniscus replacement devices. Last, due to the lack of standardization in evaluation techniques, we recommend a comprehensive battery of tests to evaluate the efficacy of meniscus replacement implants. We recommend other investigators utilize these surgical and testing techniques to establish the ovine model as the gold standard for preclinical evaluation of meniscus replacement devices.

Physical and mechanical properties of cross-linked type I collagen scaffolds derived from bovine, porcine, and ovine tendons.

Collagen scaffolds are often utilized in tissue engineering applications where their performance depends on physical and mechanical properties. This study investigated the effects of collagen source (bovine, porcine, and ovine tendon) on properties of collagen sponge scaffolds cross-linked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS). Scaffolds were tested for tensile and compressive properties, stability (resistance to enzymatic degradation), pore size, and swelling ratio. No significant differences in tensile modulus were observed, but ovine scaffolds had significantly greater ultimate strain, stress, and toughness relative to bovine and porcine scaffolds. No significant differences in compressive properties, pore size, or swelling ratio were observed as a function of collagen source. Ovine scaffolds were more resistant to collagenase degradation compared to bovine samples, which were more resistant than porcine scaffolds. In comparison to bovine scaffolds, ovine scaffolds performed equivalently or superiorly in all evaluations, and porcine scaffolds were equivalent in all properties except enzymatic stability. These results suggest that collagen sponges derived from bovine, porcine, and ovine tendon have similar physical and mechanical properties, and are all potentially suitable materials for various tissue engineering applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2685-2692, 2016.