Scaffolds For Soft Tissue Regeneration Research Articles

Inversely engineered biomimetic flexible network scaffolds for soft tissue regeneration.

Graft-host mechanical mismatch has been a longstanding issue in clinical applications of synthetic scaffolds for soft tissue regeneration. Although numerous efforts have been devoted to resolve this grand challenge, the regenerative performance of existing synthetic scaffolds remains limited by slow tissue growth (comparing to autograft) and mechanical failures. We demonstrate a class of rationally designed flexible network scaffolds that can precisely replicate nonlinear mechanical responses of soft tissues and enhance tissue regeneration via reduced graft-host mechanical mismatch. Such flexible network scaffold includes a tubular network frame containing inversely engineered curved microstructures to produce desired mechanical properties, with an electrospun ultrathin film wrapped around the network to offer a proper microenvironment for cell growth. Using rat models with sciatic nerve defects or Achilles tendon injuries, our network scaffolds show regenerative performances evidently superior to that of clinically approved electrospun conduit scaffolds and achieve similar outcomes to autologous nerve transplantation in prevention of target organ atrophy and recovery of static sciatic index.

Cobweb-Inspired Micro/Nanostructured Scaffolds for Soft Tissue Regeneration with Inhibition Effect of Fibrosis under Dynamic Environment.

In soft tissue repair, fibrosis can lead to repair failure and long-term chronic pain in patients. Excessive mechanical stimulation of fibroblasts is one of the causes of fibrosis during abdominal wall regeneration. Inspired by the cobweb, a polycaprolactone beaded fiber is prepared by electrospinning. The cobweb-inspired structure attenuates the mechanical stimulation of cells under a dynamic environment. Nano-protrusions are introduced into the scaffold for further inhibition of fibrosis by self-induced crystallization. A machine is built for in vitro dynamic culture and rat abdominal subcutaneous embedding experiments are performed to verify the inhibiting effect of fibrosis in a dynamic environment in vivo. Results show that the expression of integrin β1 and α-smooth muscle actin is inhibited by the cobweb-inspired structure under dynamic culture. The results of hematoxylin and eosin and Masson's trichrome indicate that the cobweb-inspired structure has a good inhibitory effect on fibrosis in a dynamic environment in vivo. In general, the cobweb-inspired scaffold with nano-protrusions has a good ability to inhibit fibrosis under both static and dynamic environments. It is believed that the scaffold has promising applications in the field of inhibiting fibrosis caused by mechanical stimulation.

Femtosecond laser-made 3D micro-chainmail scaffolds towards regenerative medicine

Regenerative medicine is a rapidly developing field with far-reaching potential. To sustain progress in this field, new advanced structures are needed. These include scaffolds for cell growth. Direct laser writing (DLW) based on femtosecond laser multi-photon polymerization (MPP) was shown to be an attractive technology for such structure fabrication as it combines vast selection of materials and the possibility to choose feature size in scaffold to be smaller, bigger, or around the same size as a cell. At the same, there are issues related to throughput which limit the widespread implementation of MPP for scaffold manufacturing. It is further compounded by some material limitations making it difficult to print mechanically flexible scaffolds for soft tissue regeneration. In this paper, we propose printing mechanically flexible scaffolds out of mechanically rigid material SZ2080 by employing chainmail architecture. We explore capabilities to optimize the printing procedure of this kind of scaffold, achieving printing times of less than 10 min for a 1 × 1 mm scaffold while maintaining micro-level precision. The superb biocompatibility of such scaffolds is shown both qualitatively and quantitatively and is proven to be independent of the used photoinitiator. Finally, manipulations of scaffolds with cells are performed with no adverse impact on the cell viability or proliferation after such procedures. Overall, this work proposes a methodology for rapid printing of shape-shifting scaffolds which could be used in regenerative medicine both for cell cultivation and potential direct implantation into soft tissue.

Porous cellulose-collagen scaffolds for soft tissue regeneration: influence of cellulose derivatives on mechanical properties and compatibility with adipose-derived stem cells

This study compares two types of bioresorptive cellulose, i.e., calcium-sodium salt of oxidized cellulose (OC) and sodium salt of carboxymethylcellulose (CMC). It investigates which type would be preferable as an implant material in terms of biocompatibility, biomechanical and biological properties, and also in terms of its behavior in combination with collagen fibrils (Col) in composite Col/OC or Col/CMC scaffolds. OC significantly supported the stiffness and elasticity of Col fibrils, whereas CMC significantly reduced these properties. OC also enabled a strong interaction with Col fibrils even in a moist environment, accompanied by a significant drop in elastic modulus. The addition of cellulose did not significantly influence scaffold porosity; however, changes in surface morphology and the lower swelling capacity of OC, with a degree of oxidation of its chains between 16 and 24%, supported the idea of improved cell-material interaction. The elasticity and the stiffness of Col/OC guided human adipose-derived stem cells (hADSCs) to significantly higher adhesion, proliferation, and metabolic activity. On the contrary, the Col/CMC provided only limited mechanical support for the cells and inhibited their attachment and proliferation, although without any signs of cytotoxicity. This phenomenon could be used for future control of the differentiation of hADSCs towards a desired phenotype to generate advanced tissue replacements using modern methods of tissue engineering. The oxidation of cellulose resulted in a firmer scaffolding material, as required in vascular or skin tissue engineering. CMC is suitable for moist wound healing, e.g. as a mucoadhesive gel, where cell adhesion is not desirable.Graphical abstract

Anisotropic hair keratin‐dopamine composite scaffolds exhibit strain‐stiffening properties

Human hair keratin (HHK) has been successfully explored as raw materials for three-dimensional scaffolds for soft tissue regeneration due to its excellent biocompatibility and bioactivity. However, none of the reported HHK based scaffolds is able to replicate the strain-stiffening capacity of living tissues when responding to large deformations. In the present study, strain-stiffening property was achieved in scaffolds fabricated from HHK via a synergistic effect of well-defined, aligned microstructure and chemical crosslinking. Directed ice-templating method was used to fabricate HHK-based scaffolds with highly aligned (anisotropic) microstructure while oxidized dopamine (ODA) was used to crosslink covalently to HHKs. The resultant HHK-ODA scaffolds exhibited strain-stiffening behavior characterized by the increased gradient of the stress-strain curve after the yield point. Both ultimate tensile strength and the elongation at break were enhanced significantly (~700 kPa, ~170%) in comparison to that of HHK scaffolds lacking of aligned microstructure or ODA crosslinking. In vitro cell culture studies indicated that HHK-ODA scaffolds successfully supported human dermal fibroblasts (HDFs) adhesion, spreading and proliferation. Moreover, anisotropic HHK-ODA scaffolds guided cell growth in alignment with the defined microstructure as shown by the highly organized cytoskeletal networks and nuclei distribution. The findings suggest that HHK-ODA scaffolds, with strain-stiffening properties, biocompatibility and bioactivity, have the potential to be applied as biomimetic matrices for soft tissue regeneration.

Characterization of Anisotropic Human Hair Keratin Scaffolds Fabricated via Directed Ice Templating.

Human hair keratin (HHK) is successfully exploited as raw materials for 3D scaffolds for soft tissue regeneration owing to its excellent biocompatibility and bioactivity. However, most HHK scaffolds are not able to achieve the anisotropic mechanical properties of soft tissues such as tendons and ligaments due to lack of tunable, well-defined microstructures. In this study, directed ice templating method is used to fabricate anisotropic HHK scaffolds that are characterized by aligned pores (channels) in between keratin layers in the longitudinal plane. In contrast, pores in the transverse plane maintain a homogenous rounded morphology. Channel widths throughout the scaffolds range from ≈5 to ≈15µm and are tunable by varying the freezing temperature. In comparison with HHK scaffolds with random, isotropic pore structures, the tensile strength of anisotropic HHK scaffolds is enhanced significantly by up to fourfolds (≈200 to ≈800kPa) when the tensile load is applied in the direction parallel to the aligned pores. In vitro results demonstrate that the anisotropic HHK scaffolds are able to support human dermal fibroblast adhesion, spreading, and proliferation. The findings suggest that HHK scaffolds with well-defined, aligned microstructure hold promise as templates for soft tissues regeneration by mimicking their anisotropic properties.

Rapid prototyping fabrication of soft and oriented polyester scaffolds for axonal guidance

Biodegradable polyesters have been extensively used for preparation of nerve guidance scaffolds, due to their high biocompatibility and defined degradation periods. However, conventional methods for fabrication of porous polyester scaffolds provide limited control over shape and micro-architecture. Here, a fabrication procedure based on 3D printing was developed to generate highly ordered and anatomically personalized, polyester scaffolds for soft tissue regeneration. Scaffolds composed of Poly-lactic-glycolic acid (PLGA) and poly-L-lactic acid (PLLA) were specifically customized for nerve injuries. This was obtained by using an oriented multi-layer printing pattern which established a linear structure in the fabricated scaffolds to match the aligned topography of nerve tissues. The oriented scaffold was shown to guide regenerating axons to linear conformations and support growth of induced pluripotent stem cell-derived neurons in vitro and in vivo in a model of spinal cord injury. The described scaffolds may advance the field of nerve regeneration. Furthermore, modifications could be integrated to generate soft implants for various types of tissues.

Adipose derived stem cells and platelet rich plasma improve the tissue integration and angiogenesis of biodegradable scaffolds for soft tissue regeneration

Current surgical reconstruction for soft tissue replacement involves lipotransfer to restore soft tissue replacements but is limited by survival and longevity of the fat tissue. Alternative approaches to overcome these limitations include using biodegradable scaffolds with stem cells with growth factors to generate soft tissue. Adipose derived stem cells (ADSCs) offer great potential to differentiate into adipose, and can be delivered using biodegradable scaffolds. However, the optimal scaffold to maximise this approach is unknown. This study investigates the biocompatibility of nanocomposite scaffolds (POSS-PCL) to deliver ADSCs with and without the addition of growth factors using platelet rich plasma (PRP) in vivo. Rat ADSCs were isolated and then seeded on biodegradable scaffolds (POSS-PCL). In addition, donor rats were used to isolate PRP to modify the scaffolds. The implants were then subcutaneously implanted for 3-months to assess the effect of PRP and ADSC on POSS-PCL scaffolds biocompatibility. Histology after explanation was examined to assess tissue integration (H&E) and collagen production (Massons Trichome). Immunohistochemistry was used to assess angiogenesis (CD3, α-SMA), immune response (CD45, CD68) and adipose formation (PPAR-γ). At 3-months PRP-ADSC-POSS-PCL scaffolds demonstrated significantly increased tissue integration and angiogenesis compared to PRP, ADSC and unmodified scaffolds (p < 0.05). In addition, PRP-ADSC-POSS-PCL scaffolds showed similar levels of CD45 and CD68 staining compared to unmodified scaffolds. Furthermore, there was increased PPAR-γ staining demonstrated at 3-months with PRP-ADSC-POSS-PCL scaffolds (p < 0.05). POSS-PCL nanocomposite scaffolds provide an effective delivery system for ADSCs. PRP and ADSC work synergistically to enhance the biocompatibility of POSS-PCL scaffolds and provide a platform technology for soft tissue regeneration.

Skin Regeneration with a Scaffold of Predefined Shape and Bioactive Peptide Hydrogels.

We developed a highly elastic customized scaffold for soft tissue regeneration and combined them with bioactive hydrogels with stem cell-inducing ability. This was done to mimic mechanical properties of native soft tissues and improve the viability of transplanted cells as well as efficiency of tissue regeneration. The proposed study was aimed at evaluating various characteristics of scaffolds and investigating their tissue-regenerating ability. Finger-shaped porous scaffolds were successfully fabricated by an indirect 3D printing of poly (L-lactide-co-ɛ-caprolactone) (PLCL), which provides high elasticity for soft tissue engineering. In addition, a self-assembling peptide hydrogel coupled with substance P (RARADADARARADADA/RARADADARARADADA-substance P, RADA16/RADA16-SP) was used to accelerate angiogenesis and recruit intrinsic mesenchymal stem cells (MSCs). This study included three kinds of groups: Group I = PLCL scaffold with human dermal fibroblasts (HDFs) (P+C), Group II = PLCL scaffold with HDFs and RADA16 (P+C+R), and Group III = PLCL scaffold with HDFs, RADA16, and RADA16-SP (P+C+R+S). The samples were implanted into immunodeficient mice subcutaneously and harvested at 1 and 4 weeks. Tissue regeneration was evaluated by histological analysis with hematoxylin and eosin (H&E) and Masson's trichrome (MT) staining. The images showed that a large number of cells were recruited into the scaffolds, and collagen was deposited in the constructs of the P+C+R+S group. Additionally, recruitment of MSCs, angiogenesis, and collagen were observed by immunofluorescence staining. The results show that the P+C+R+S group had more type I and type III collagen, which are formed in soft tissues, and were deposited on the scaffold compared with the other groups. Moreover, more blood vessels and MSCs were induced in the P+C+R+S group than in those of the P + C and P+C+R groups. Consequently, the results suggest that the construct of the customized porous PLCL scaffold and RADA16/RADA16-SP hydrogel could be a good treatment modality to treat skin defects.

Exploring the potential of polyurethane-based soft foam as cell-free scaffold for soft tissue regeneration

Reconstructive treatment after trauma and tumor resection would greatly benefit from an effective soft tissue regeneration. The use of cell-free scaffolds for adipose tissue regeneration in vivo is emerging as an attractive alternative to tissue-engineered constructs, since this approach avoids complications due to cell manipulation and lack of synchronous vascularization. In this study, we developed a biodegradable polyurethane-based scaffold for soft tissue regeneration, characterized by an exceptional combination between softness and resilience. Exploring the potential as a cell-free scaffold required profound understanding of the impact of its intrinsic physico-chemical properties on the biological performance in vivo. We investigated the effect of the scaffold’s hydrophilic character, degradation kinetics, and internal morphology on (i) the local inflammatory response and activation of MGCs (foreign body response); (ii) its ability to promote rapid vascularisation, cell infiltration and migration through the scaffold over time; and (iii) the grade of maturation of the newly formed tissue into vascularized soft tissue in a murine model. The study revealed that soft tissue regeneration in vivo proceeded by gradual infiltration of undifferentiated mesenchymal cells though the periphery toward the center of the scaffold, where the rapid formation of a functional and well-formed vascular network supported cell viability overtime. Statement of significanceExploring the potential of polyurethane-based soft foam as cell-free scaffold for soft tissue regeneration.In this work, we address the unmet need for synthetic functional soft tissue substitutes that provide adequate biological and mechanical support to soft tissue. We developed a series of flexible cross-linked polyurethane copolymer scaffolds with remarkable fatigue-resistance and tunable physico-chemical properties for soft tissue regeneration in vivo. Accordingly, we could extend the potential of this class of biomaterials, which was so far confined for bone and osteochondral tissue regeneration, to other types of connective tissue.

Decellularized Adipose Tissue Scaffolds for Soft Tissue Regeneration and Adipose-Derived Stem/Stromal Cell Delivery.

Surgically discarded adipose tissue is not only an abundant source of multipotent adipose-derived stem/stromal cells (ASCs) but can also be decellularized to obtain a biomimetic microenvironment for tissue engineering applications. The decellularization methods involve processing excised fat through a series of chemical, mechanical, and enzymatic treatment stages designed to extract cells, cellular components, and lipid from the tissues. This process yields a complex 3D bioscaffold enriched in collagens that mimics the biochemical and biomechanical properties of the native extracellular matrix (ECM). For ASC culture and delivery, decellularized adipose tissue (DAT) provides a cell-supportive platform that is conducive to adipogenesis. While DAT can be applied in its intact form as an off-the-shelf adipogenic matrix, it can also be used as an ECM source for the fabrication of an array of other scaffold formats including adipose ECM-derived microcarriers and porous foams. In this chapter, we describe the methods developed in our lab to decellularize human adipose tissue and to further process it into a variety of scaffolding materials for a range of applications in soft tissue regeneration, wound healing, and cell culture.

Collagen-Fibrinogen Lyophilised Scaffolds for Soft Tissue Regeneration.

A significant body of research has considered collagen as a scaffold material for soft tissue regeneration. The main structural component of extra-cellular matrix (ECM), collagen’s advantages over synthetic polymers are numerous. However, for applications where higher stiffness and stability are required, significant cross-linking may affect bioactivity. A carbodiimide (EDC) cross-linking route consumes carboxylate groups that are key to collagen’s essential cell recognition motifs (GxOGER). Fibrinogen was considered as a promising additive as it plays a key role in the process of wound repair and contains RGD integrin binding sites which bind to a variety of cells, growth factors and cytokines. Fibrinogen’s binding sites however, also contain the same carboxylate groups as collagen. We have successfully produced highly interconnected, porous collagen-fibrinogen scaffolds using a lyophilisation technique and micro-computed tomography demonstrated minimal influence of either fibrinogen content or cross-linking concentration on the scaffold structure. The specific biological effect of fibrinogen additions into cross-linked collagen are considered by using films as a model for the struts of bulk scaffolds. By considering various additions of fibrinogen to the collagen film with increasing degrees of cross-linking, this study demonstrates a significant biological advantage with fibrinogen addition across the cross-linking concentrations typically applied to collagen-based scaffolds.

Hybrid hydrogels based on keratin and alginate for tissue engineering.

Novel hybrid hydrogels based on alginate and keratin were successfully produced for the first time. The self-assembly properties of keratin, and its ability to mimic the extracellular matrix were combined with the excellent chemical and mechanical stability and biocompatibility of alginate to produce 2D and 3D hybrid hydrogels. These hybrid hydrogels were prepared using two different approaches: sonication, to obtain 2D hydrogels, and a pressure-driven extrusion technique to produce 3D hydrogels. All results indicated that the composition of the hydrogels had a significant effect on their physical properties, and that they can easily be tuned to obtain materials suitable for biological applications. The cell-material interaction was assessed through the use of human umbilical vein endothelial cells, and the results demonstrated that the alginate/keratin hybrid biomaterials supported cell attachment, spreading and proliferation. The results proved that such novel hybrid hydrogels might find applications as scaffolds for soft tissue regeneration.

Injectable polyMIPE scaffolds for soft tissue regeneration

Injury caused by trauma, burns, surgery, or disease often results in soft tissue loss leading to impaired function and permanent disfiguration. Tissue engineering aims to overcome the lack of viable donor tissue by fabricating synthetic scaffolds with the requisite properties and bioactive cues to regenerate these tissues. Biomaterial scaffolds designed to match soft tissue modulus and strength should also retain the elastomeric and fatigue-resistant properties of the tissue. Of particular design importance is the interconnected porous structure of the scaffold needed to support tissue growth by facilitating mass transport. Adequate mass transport is especially true for newly implanted scaffolds that lack vasculature to provide nutrient flux. Common scaffold fabrication strategies often utilize toxic solvents and high temperatures or pressures to achieve the desired porosity. In this study, a polymerized medium internal phase emulsion (polyMIPE) is used to generate an injectable graft that cures to a porous foam at body temperature without toxic solvents. These poly(ester urethane urea) scaffolds possess elastomeric properties with tunable compressive moduli (20–200 kPa) and strengths (4–60 kPa) as well as high recovery after the first conditioning cycle (97–99%). The resultant pore architecture was highly interconnected with large voids (0.5–2 mm) from carbon dioxide generation surrounded by water-templated pores (50–300 μm). The ability to modulate both scaffold pore architecture and mechanical properties by altering emulsion chemistry was demonstrated. Permeability and form factor were experimentally measured to determine the effects of polyMIPE composition on pore interconnectivity. Finally, initial human mesenchymal stem cell (hMSC) cytocompatibility testing supported the use of these candidate scaffolds in regenerative applications. Overall, these injectable polyMIPE foams show strong promise as a biomaterial scaffold for soft tissue repair.

Hyaluronic acid-recombinant gelatin gels as a scaffold for soft tissue regeneration

An array of different types of hyaluronic acid (HA)- and collagen-based products is available for filling soft-tissue defects. A major drawback of the current soft-tissue fillers is their inability to induce cell infiltration and new tissue formation. Our aim is to develop novel biodegradable injectable gels which induce soft tissue regeneration, initially resulting in integration and finally replacement of the gel with new autologous tissue. Two reference gels of pure HA, monophasic HA-1 and micronised HA-2, were used. Furthermore, both gels were mixed with recombinant gelatin (RG) resulting in HA-1+RG and HA-2+RG. All gels were subcutaneously injected on the back of rats and explanted after 4 weeks. Addition of RG to HA-1 resulted in stroma formation (neovascularisation and ECM deposition) which was restricted to the outer rim of the HA-1+RG gel. In contrast, addition of RG to HA-2 induced stroma formation throughout the gel. The RG component of the gel was degraded by macrophages and giant cells and subsequently replaced by new vascularised tissue. Immunohistochemical staining showed that the extracellular matrix components collagen I and III were deposited throughout the gel. In conclusion, this study shows the proof of principle that addition of RG to HA-2 results in a novel injectable gel capable of inducing soft tissue regeneration. In this gel HA has a scaffold function whereas the RG component induces new tissue formation, resulting in proper vascularisation and integration of the HA-2+RG gel with the autologous tissue.

Degradation‐induced changes of mechanical properties of an electro‐spun polyester‐urethane scaffold for soft tissue regeneration

The aim of this study was the in vitro investigation of the change in mechanical properties of a fast-degrading electro-spun polymeric scaffold for the use in soft tissue regenerative implants. Tubular scaffolds were electro-spun from a DegraPol® D30 polyesther-urethane solution (target outer diameter: 5.0 mm; scaffold wall thickness: 0.99 ± 0.18 mm). Scaffold samples were subjected to hydrolytic in vitro degradation for up to 34 days. The fiber network structure and fiber surfaces were inspected on scanning electron micrographs. Following vacuum drying and determination of mass, flat samples (9.69 ± 0.21 × 18.47 ± 2.62 mm, n = 5) underwent uni-axial tensile testing (5 load cycles, strain ε = 0 to 20%; final extension to failure) in circumferential scaffold direction after 5, 10, 14, 18, 22, 26, 30, and 34 days of degradation. Scaffold mass did not change with degradation. Maximum elastic modulus, maximum stress and associated strain were E(max) = 1.14 ± 0.23 MPa, σ(max) = 0.52 ± 0.12 MPa and ε(max) = 176.8 ± 21.9% before degradation and E(max) = 0.43 ± 0.26 MPa, σ(max) = 0.033 ± 0.028 MPa and ε(max) = 24.6 ± 3.0% after 34 days of degradation. The deterioration of mechanical properties was not reflected in the ultrastructural surface morphology of the fibers. The current exploratory study provides a basis for the development of constitutive computational models of biodegradable scaffolds with future extension of the investigation most importantly to capture mechanical effects of regenerating tissue. Future studies will include degradation in biological fluids and assessment of molecular weight for an advanced understanding of the material changes during degradation.

Enzymatic degradation of the hydrogels based on synthetic poly(α-amino acid)s

Biodegradable hydrogels are studied as potential scaffolds for soft tissue regeneration. In this work biodegradable hydrogels were prepared from synthetic poly(α-amino acid)s, poly(AA)s. The covalently crosslinked gels were formed by radical copolymerization of methacryloylated poly(AA)s, e.g. poly[N (5)-(2-hydroxy-ethyl)-L-glutamine-ran-L-alanine-ran-N (6)-methacryloyl-L-lysine], as a multifunctional macro-monomer with a low-molecular-weight methacrylic monofunctional monomer, e.g. 2-hydroxyethyl methacrylate (HEMA). Methacryloylated copolypeptides were synthesized by polymerization of N-carboxyanhydrides of respective amino acids and subsequent side-chain modification. Due to their polypeptide backbone, synthetic poly(AA)s are cleavable in biological environment by enzyme-catalyzed hydrolysis. The feasibility of enzymatic degradation of poly(AA)s alone and the hydrogels made from them was studied using elastase, a matrix proteinase involved in tissue healing processes, as a model enzyme. Specificity of elastase for cleavage of polypeptide chains behind the L-alanine residues was reflected in faster degradation of L-alanine-containing copolymers as well as of hydrogels composed of them.

Impacts of stereoregularity and stereocomplex formation on physicochemical, protein adsorption and cell adhesion behaviors of star-shaped 8-arms poly(ethylene glycol)–poly(lactide) block copolymer films

Biodegradable stereocomplex film exhibiting soft and stretchy character was prepared by simply blending between enantiomeric 8-arms poly(ethylene glycol)- block-poly( l-lactide) (8-arms PEG- b-PLLA) and 8-arms PEG- b-PDLA copolymers with star-shaped structure. The stereocomplex film exhibited higher T g and PLA crystallinity than those of original copolymer films. Effects of stereoregularity and stereocomplexation on protein adsorption and L929 cells attachment/proliferation behaviors onto the films were analyzed from the viewpoint to design a new class of implantable soft biomaterial. The stereocomplex film was found to exhibit large amount of protein adsorption than original films. Furthermore, cell attachment efficiency and proliferation rate on the film were significantly enhanced by stereocomplexation. This stereocomplex material is expected to be applicable as degradable temporary scaffold for soft tissue regeneration. Consequently, it was indicated that the stereocomplex formation could be proposed to be a novel method to control the protein- and cell-adhesive properties of biodegradable matrix composed of PEG–PLA copolymer.

Characterizing fibroblast migration on discrete collagen threads for applications in tissue regeneration.

Collagen threads with mechanical properties and fibrillar substructure similar to native tissue have been synthesized for the repair of injured tendon and ligament. While these scaffolding materials have demonstrated the potential for inducing tissue regeneration, one limitation has been an insufficient rate of tissue ingrowth for complete regeneration. We hypothesize that the structural hierarchy and biochemical cues on the surfaces of these threads will enhance the rate of cell migration and ultimately the rate of new tissue ingrowth. We developed an in vitro assay to measure the effects of various collagen sources and crosslinking on the rate of fibroblast migration on the surfaces of collagen threads. Threads were suspended from elevated platforms and seeded with fibroblast-populated collagen lattices. Cell migration rates ranging from 0.75 to 1.25 mm/day were measured as the fibroblasts left the lattices and migrated onto various thread types. Threads self-assembled from type I collagen were found to have migration rates similar to native tendon threads while crosslinking by severe dehydration decreased the rate. This novel in vitro model system allows examination of cell migration from a wound margin onto biomaterials to determine the effects of various cell types, matrix materials, and surface biochemistries on cell-matrix interactions. Ultimately, this assay will allow us to identify design parameters that will be most effective for enhancing the rate of tissue ingrowth on fiber-based collagen scaffolds for soft tissue regeneration.

Fibroblast culture on surface-modified poly (glycolide-co-ε-caprolactone) scaffold for soft tissue regeneration

Novel porous matrices made of a copolymer of glycolide (G) and ε-caprolactone (CL) (51 : 49, Mw 103 000) was prepared for tissue engineering using a solvent-casting particulate leaching method. Poly(glycolide-co-ε-caprolactone) (PGCL) copolymer showed a rubber-like elastic characteristic, in addition to an amorphous property and fast biodegradability. In order to investigate the effect on the fibroblast culture, PGCL scaffolds of varying porosity and pore size, in addition to surfacehydrolysis or collagen coating, were studied. The large pore-sized scaffold (pore size >150 μm) demonstrated a much greater cell adhesion and proliferation than the small pore-sized one. In addition, the higher porosity, the better the cell adhesion and proliferation. The surface-hydrolyzed PGCL scaffold showed enhanced cell adhesion and proliferation compared with the unmodified one. Type I collagen coating revealed a more pronounced contribution for increased cell interactions than the surface-hydrolyzed one. These results demonstrate that surface-modified PGCL scaffold can provide a suitable substrate for fibroblast culture, especially in the case of soft tissue regenerations.