Evaluation of a self-fitting, shape memory polymer scaffold in a rabbit calvarial defect model.

Self-fitting scaffolds prepared from biodegradable poly(ε-caprolactone)-diacrylate (PCL-DA) have been developed for the treatment of craniomaxillofacial (CMF) bone defects. As a thermoresponsive shape memory polymer (SMP), with the mere exposure to warm saline, these porous scaffolds achieve a conformal fit in defects. This behavior was expected to be advantageous to osseointegration and thus bone healing. Herein, for an initial assessment of their regenerative potential, a pilot in vivo study was performed using a rabbit calvarial defect model. Exogenous growth factors and cells were excluded from the scaffolds. Key scaffold material properties were confirmed to be maintained following gamma sterilization. To assess scaffold integration and neotissue infiltration along the defect perimeter, non-critically sized (d = 8 mm) bilateral calvarial defects were created in 12 New Zealand white rabbits. Bone formation was assessed at 4 and 16 weeks using histological analysis and micro-CT, comparing defects treated with an SMP scaffold (d = 9 mm x t = 1 or 2 mm) to untreated defects (i.e. defects able to heal without intervention). To further assess osseointegration, push-out tests were performed at 16 weeks and compared to defects treated with poly(ether ether ketone) (PEEK) discs (d = 8.5 mm x t = 2 mm). The results of this study confirmed that the SMP scaffolds were biocompatible and highly conducive to bone formation and ingrowth at the perimeter. Ultimately, this resulted in similar bone volume and surface area versus untreated defects and superior performance in push-out testing versus defects treated with PEEK discs. STATEMENT OF SIGNIFICANCE: Current treatments of craniomaxillofacial (CMF) bone defects include biologic and synthetic grafts but they are limited in their ability to form good contact with adjacent tissue. A regenerative engineering approach using a biologic-free scaffold able to achieve conformal fitting represents a potential "off-the-shelf" surgical product to heal CMF bone defects. Having not yet been evaluated in vivo, this study provided the preliminary assessment of the bone healing potential of self-fitting PCL scaffolds using a rabbit calvarial defect model. The study was designed to assess scaffold biocompatibility as well as bone formation and ingrowth using histology, micro-CT, and biomechanical push-out tests. The favorable results provide a basis to pursue establishing self-fitting scaffolds as a treatment option for CMF defects.

Smart scaffolds: shape memory polymers (SMPs) in tissue engineering.

Smart scaffolds based on shape memory polymer (SMPs) have been increasingly studied in tissue engineering. The unique shape actuating ability of SMP scaffolds has been utilized to improve delivery and/or tissue defect filling. In this regard, these scaffolds may be self-deploying, self-expanding, or self-fitting. Smart scaffolds are generally thermoresponsive or hydroresponsive wherein shape recovery is driven by an increase in temperature or by hydration, respectively. Most smart scaffolds have been directed towards regenerating bone, cartilage, and cardiovascular tissues. A vast variety of smart scaffolds can be prepared with properties targeted for a specific tissue application. This breadth of smart scaffolds stems from the variety of compositions employed as well as the numerous methods used to fabricated scaffolds with the desired morphology. Smart scaffold compositions span across several distinct classes of SMPs, affording further tunability of properties using numerous approaches. Specifically, these SMPs include those based on physically cross-linked and chemically cross-linked networks and include widely studied shape memory polyurethanes (SMPUs). Various additives, ranging from nanoparticles to biologicals, have also been included to impart unique functionality to smart scaffolds. Thus, given their unique functionality and breadth of tunable properties, smart scaffolds have tremendous potential in tissue engineering.

Shape memory polymer (SMP) scaffolds with improved self-fitting properties.

"Self-fitting" shape memory polymer (SMP) scaffolds prepared as semi-interpenetrating networks (semi-IPNs) with crosslinked linear-poly(ε-caprolactone)-diacrylate (PCL-DA, Mn∼10 kg mol-1) and linear-poly(l-lactic acid) (PLLA, Mn∼15 kg mol-1) [75/25 wt%] exhibited robust mechanical properties and accelerated degradation rates versus a PCL-DA scaffold control. However, their potential to treat irregular craniomaxillofacial (CMF) bone defects is limited by their relatively high fitting temperature (Tfit∼55 °C; related to the Tm of PCL) required for shape recovery (i.e. expansion) and subsequent shape fixation during press fitting of the scaffold, which can be harmful to surrounding tissue. Additionally, the viscosity of the solvent-based precursor solutions, cast over a fused salt template during fabrication, can limit scaffold size. Thus, in this work, analogous semi-IPN SMP scaffolds were formed with a 4-arm star-PCL-tetracryalate (star-PCL-TA) (Mn∼10 kg mol-1) and star-PLLA (Mn∼15 kg mol-1). To assess the impact of a star-polymer architecture, four semi-IPN compositions were prepared: linear-PCL-DA/linear-PLLA (L/L), linear-PCL-DA/star-PLLA (L/S), star-PCL-TA/linear-PLLA (S/L) and star-PCL-TA/star-PLLA (S/S). Two PCL controls were also prepared: LPCL (i.e. 100% linear-PCL-DA) and SPCL (i.e. 100% star-PCL-TA). The S/S semi-IPN scaffold exhibited particularly desirable properties. In addition to achieving a lower, tissue-safe Tfit (∼45 °C), it exhibited the fastest rate of degradation which is anticipated to more favourably permit neotissue infiltration. The radial expansion pressure exerted by the S/S semi-IPN scaffold at Tfit was greater than that of LPCL, which is expected to enhance osseointegration and mechanical stability. The intrinsic viscosity of the S/S semi-IPN macromer solution was also reduced such that larger scaffold specimens could be prepared.

Intrinsic osteoinductivity of PCL-DA/PLLA semi-IPN shape memory polymer scaffolds.

Engineering osteoinductive, self-fitting scaffolds offers a potential treatment modality to repair irregularly shaped craniomaxillofacial bone defects. Recently, we innovated on osteoinductive poly(ε-caprolactone)-diacrylate (PCL-DA) shape memory polymers (SMPs) to incorporate poly-L-lactic acid (PLLA) into the PCL-DA network, forming a semi-interpenetrating network (semi-IPN). Scaffolds formed from these PCL-DA/PLLA semi-IPNs display stiffnesses within the range of trabecular bone and accelerated degradation relative to scaffolds formed from slowly degrading PCL-DA SMPs. Herein, we demonstrate for the first time that PCL-DA/PLLA semi-IPN SMP scaffolds show increased intrinsic osteoinductivity relative to PCL-DA. We also confirm that application of a bioinspired polydopamine (PD) coating further improves the osteoinductive capacity of these PCL-DA/PLLA semi-IPN SMPs. In the absence of osteogenic supplements, protein level assessment of human mesenchymal stem cells (h-MSCs) cultured in PCL-DA/PLLA scaffolds revealed an increase in expression of osteogenic markers osterix, bone morphogenetic protein-4 (BMP-4), and collagen 1 alpha 1 (COL1A1), relative to PCL-DA scaffolds and osteogenic medium controls. Likewise, the expression of runt-related transcription factor 2 (RUNX2) and BMP-4 was elevated in the presence of PD-coating. In contrast, the chondrogenic and adipogenic responses associated with the scaffolds matched or were reduced relative to osteogenic medium controls, indicating that the scaffolds display intrinsic osteoinductivity.

PCL-Based Shape Memory Polymer Semi-IPNs: The Role of Miscibility in Tuning the Degradation Rate.

The utility of poly(ε-caprolactone) (PCL) as a shape memory polymer (SMP) may be improved by accelerating its degradation. Recently, we have reported novel semi-interpenetrating networks (semi-IPNs) composed of cross-linked PCL diacrylate (PCL-DA) and thermoplastic poly(l-lactic acid) (PLLA) that exhibited SMP behavior, accelerated degradation, and enhanced moduli versus the PCL-DA control. Herein, we systematically varied the thermoplastic component of the PCL-based semi-IPNs, incorporating homo- and copolymers based on lactic acid of different Mn, hydrophilicity, and crystallinity. Specifically, semicrystalline PLLAs of different Mns (7.5, 15, 30, and 120 kDa) were explored as the thermoplastics in the semi-IPNs. Additionally, to probe crystallinity and hydrophilicity, amorphous (or nearly amorphous) thermoplastics of different hydrophilicities (PDLLA and PLGAs 85:15, 70:30, and 50:50, l-lactide:glycolide mole % ratio) were employed. For all semi-IPNs, the wt % ratio of the cross-linked PCL-DA to thermoplastic was 75:25. The nature of the thermoplastics was linked to semi-IPN miscibility and the trends in accelerated degradation rates.