Biomaterials-Based Regenerative Strategies for Skin Tissue Wound Healing.

Skin tissue wound healing proceeds through four major stages, including hematoma formation, inflammation, and neo-tissue formation, and culminates with tissue remodeling. These four steps significantly overlap with each other and are aided by various factors such as cells, cytokines (both anti- and pro-inflammatory), and growth factors that aid in the neo-tissue formation. In all these stages, advanced biomaterials provide several functional advantages, such as removing wound exudates, providing cover, transporting oxygen to the wound site, and preventing infection from microbes. In addition, advanced biomaterials serve as vehicles to carry proteins/drug molecules/growth factors and/or antimicrobial agents to the target wound site. In this review, we report recent advancements in biomaterials-based regenerative strategies that augment the skin tissue wound healing process. In conjunction with other medical sciences, designing nanoengineered biomaterials is gaining significant attention for providing numerous functionalities to trigger wound repair. In this regard, we highlight the advent of nanomaterial-based constructs for wound healing, especially those that are being evaluated in clinical settings. Herein, we also emphasize the competence and versatility of the three-dimensional (3D) bioprinting technique for advanced wound management. Finally, we discuss the challenges and clinical perspective of various biomaterial-based wound dressings, along with prospective future directions. With regenerative strategies that utilize a cocktail of cell sources, antimicrobial agents, drugs, and/or growth factors, it is expected that significant patient-specific strategies will be developed in the near future, resulting in complete wound healing with no scar tissue formation.

Enhancing the Surface Properties of a Bioengineered Anterior Cruciate Ligament Matrix for Use with Point-of-Care Stem Cell Therapy.

We have previously developed a poly(L-lactic) acid (PLLA) bioengineered anterior cruciate ligament (ACL) matrix that has demonstrated enhanced healing when seeded with primary ACL cells prior to implantation in a rabbit model, as compared with the matrix alone. This suggests that improving cell adhesion on the matrix may beneficially affect the healing response and long-term performance of the bioengineered ACL matrix. One regenerative engineering approach involves enhancing the surface properties of the matrix to support cell adhesion and growth in combination with point-of-care stem cell therapy. Herein, we studied the cell adhesion properties of PLLA braided microfiber matrices enhanced through the physical adsorption of fibronectin and air plasma treatment. We evaluated the kinetics and binding efficiency of fibronectin onto matrices at three time points and three fibronectin concentrations. Incubating the matrix for 120 min in a solution of 25 mg mL-1 fibronectin achieved the greatest binding efficiency to the matrix and cellular adhesion. Exposing the matrices to air plasma treatment for 5 min before fibronectin adsorption significantly enhanced the cell adhesion of rabbit bone marrow-derived mesenchymal stem cells (R-BMMSCs) 24 h post cell seeding. Finally, cellular proliferation was monitored for up to 21 d, the matrices were exposed to air plasma treatment, and fibronectin adsorption was found to result in enhanced cell number. These findings suggest that exposure to air plasma treatment and fibronectin adsorption enhances the cellular adhesion of PLLA braided microfiber matrices and may improve the clinical efficacy of the matrix in combination with point-of-care stem cell therapies.

Aliphatic Polyester Nanofibers Functionalized with Cyclodextrins and Cyclodextrin-Guest Inclusion Complexes.

The fabrication of nanofibers by electrospinning has gained popularity in the past two decades; however, only in this decade, have polymeric nanofibers been functionalized using cyclodextrins (CDs) or their inclusion complexes (ICs). By combining electrospinning of polymers with free CDs, nanofibers can be fabricated that are capable of capturing small molecules, such as wound odors or environmental toxins in water and air. Likewise, combining polymers with cyclodextrin-inclusion complexes (CD-ICs), has shown promise in enhancing or controlling the delivery of small molecule guests, by minor tweaking in the technique utilized in fabricating these nanofibers, for example, by forming core⁻shell or multilayered structures and conventional electrospinning, for controlled and rapid delivery, respectively. In addition to small molecule delivery, the thermomechanical properties of the polymers can be significantly improved, as our group has shown recently, by adding non-stoichiometric inclusion complexes to the polymeric nanofibers. We recently reported and thoroughly characterized the fabrication of polypseudorotaxane (PpR) nanofibers without a polymeric carrier. These PpR nanofibers show unusual rheological and thermomechanical properties, even when the coverage of those polymer chains is relatively sparse (~3%). A key advantage of these PpR nanofibers is the presence of relatively stable hydroxyl groups on the outer surface of the nanofibers, which can subsequently be taken advantage of for bioconjugation, making them suitable for biomedical applications. Although the number of studies in this area is limited, initial results suggest significant potential for bone tissue engineering, and with additional bioconjugation in other areas of tissue engineering. In addition, the behaviors and uses of aliphatic polyester nanofibers functionalized with CDs and CD-ICs are briefly described and summarized. Based on these observations, we attempt to draw conclusions for each of these combinations, and the relationships that exist between their presence and the functional behaviors of their nanofibers.

Regenerative Engineering of the Rotator Cuff of the Shoulder.

Rotator cuff tears often heal poorly, leading to re-tears after repair. This is in part attributed to the low proliferative ability of the resident cells (tendon fibroblasts and tendon-stem cells) upon injury to the rotator cuff tissue and the low vascularity of the tendon insertion. In addition, surgical outcomes of current techniques used in clinical settings are often suboptimal, leading to the formation of neo-tissue with poor biomechanics and structural characteristics, which results in re-tears. This has prompted interest in a new approach, which we term as "Regenerative Engineering", for regenerating rotator cuff tendons. In the Regenerative Engineering paradigm, roles played by stem cells, scaffolds, growth factors/small molecules, the use of local physical forces, and morphogenesis interplayed with clinical surgery techniques may synchronously act, leading to synergistic effects and resulting in successful tissue regeneration. In this regard, various cell sources such as tendon fibroblasts and adult tissue-derived stem cells have been isolated, characterized, and investigated for regenerating rotator cuff tendons. Likewise, numerous scaffolds with varying architecture, geometry, and mechanical characteristics of biologic and synthetic origin have been developed. Furthermore, these scaffolds have been also fabricated with biochemical cues (growth factors and small molecules), facilitating tissue regeneration. In this Review, various strategies to regenerate rotator cuff tendons using stem cells, advanced materials, and factors in the setting of physical forces under the Regenerative Engineering paradigm are described.

Reorganizing Polymer Chains with Cyclodextrins.

During the past several years, we have been utilizing cyclodextrins (CDs) to nanostructure polymers into bulk samples whose chain organizations, properties, and behaviors are quite distinct from neat bulk samples obtained from their solutions and melts. We first form non-covalently bonded inclusion complexes (ICs) between CD hosts and guest polymers, where the guest chains are highly extended and separately occupy the narrow channels (~0.5⁻1.0 nm in diameter) formed by the columnar arrangement of CDs in the IC crystals. Careful removal of the host crystalline CD lattice from the polymer-CD-IC crystals leads to coalescence of the guest polymer chains into bulk samples, which we have repeatedly observed to behave distinctly from those produced from their solutions or melts. While amorphous polymers coalesced from their CD-ICs evidence significantly higher glass-transition temperatures, Tgs, polymers that crystallize generally show higher melting and crystallization temperatures (Tms, Tcs), and some-times different crystalline polymorphs, when they are coalesced from their CD-ICs. Formation of CD-ICs containing two or more guest homopolymers or with block copolymers can result in coalesced samples which exhibit intimate mixing between their common homopolymer chains or between the blocks of the copolymer. On a more practically relevant level, the distinct organizations and behaviors observed for polymer samples coalesced from their CD-ICs are found to be stable to extended annealing at temperatures above their Tgs and Tms. We believe this is a consequence of the structural organization of the crystalline polymer-CD-ICs, where the guest polymer chains included in host-IC crystals are separated and confined to occupy the narrow channels formed by the host CDs during IC crystallization. Substantial degrees of the extended and un-entangled natures of the IC-included chains are apparently retained upon coalescence, and are resistant to high temperature annealing. Following the careful removal of the host CD lattice from each randomly oriented IC crystal, the guest polymer chains now occupying a much-reduced volume may be somewhat "nematically" oriented, resulting in a collection of randomly oriented "nematic" regions of largely extended and un-entangled coalesced guest chains. The suggested randomly oriented nematic domain organization of guest polymers might explain why even at high temperatures their transformation to randomly-coiling, interpenetrated, and entangled melts might be difficult. In addition, the behaviors and uses of polymers coalesced from their CD-ICs are briefly described and summarized here, and we attempted to draw conclusions from and relationships between their behaviors and the unique chain organizations and conformations achieved upon coalescence.

Novel cellulose-collagen blend biofibers prepared from an amine/salt solvent system.

Cellulose/collagen biofibers were produced from ethylene diamine/potassium thiocyanate binary solvent system, with methanol as a coagulant. The dynamic viscosity of the solutions decreased with the gradual increase in the collagen content up to 40%. The elemental analysis showed incorporation of collagen into cellulose matrix, thereby demonstrating some degree of interaction with the cellulose matrix. The chemical and thermal analysis further revealed an intermolecular interaction between cellulose and the protein and improved thermal stability, respectively. Furthermore, the electron microscopy images mostly exhibited fibrillar morphology with no visible phase separation, indicating compatibility between the two phases. Moreover, biofibers containing higher cellulose content showed higher crystallinity, tensile, and birefringence properties of the composite fibers.

Poly (lactic acid)-based biomaterials for orthopaedic regenerative engineering.

Regenerative engineering converges tissue engineering, advanced materials science, stem cell science, and developmental biology to regenerate complex tissues such as whole limbs. Regenerative engineering scaffolds provide mechanical support and nanoscale control over architecture, topography, and biochemical cues to influence cellular outcome. In this regard, poly (lactic acid) (PLA)-based biomaterials may be considered as a gold standard for many orthopaedic regenerative engineering applications because of their versatility in fabrication, biodegradability, and compatibility with biomolecules and cells. Here we discuss recent developments in PLA-based biomaterials with respect to processability and current applications in the clinical and research settings for bone, ligament, meniscus, and cartilage regeneration.

Animal models of osteoarthritis: classification, update, and measurement of outcomes.

Osteoarthritis (OA) is one of the most commonly occurring forms of arthritis in the world today. It is a debilitating chronic illness causing pain and immense discomfort to the affected individual. Significant research is currently ongoing to understand its pathophysiology and develop successful treatment regimens based on this knowledge. Animal models have played a key role in achieving this goal. Animal models currently used to study osteoarthritis can be classified based on the etiology under investigation, primary osteoarthritis, and post-traumatic osteoarthritis, to better clarify the relationship between these models and the pathogenesis of the disease. Non-invasive animal models have shown significant promise in understanding early osteoarthritic changes. Imaging modalities play a pivotal role in understanding the pathogenesis of OA and the correlation with pain. These imaging studies would also allow in vivo surveillance of the disease as a function of time in the animal model. This review summarizes the current understanding of the disease pathogenesis, invasive and non-invasive animal models, imaging modalities, and pain assessment techniques in the animals.

Fabrication and Characterization of Poly(ε-caprolactone)/α-Cyclodextrin Pseudorotaxane Nanofibers.

Multifunctional scaffolds comprising neat poly(ε-caprolactone) (PCL) and α-cyclodextrin pseudorotaxanated in α-cyclodextrin form have been fabricated using a conventional electrospinning process. Thorough in-depth characterizations were performed on the pseudorotaxane nanofibers prepared from chloroform (CFM) and CFM/dimethylformamide (DMF) utilizing scanning electron microscopy (SEM), transmission electron microscopy (TEM), rheology, differential scanning calorimetry (DSC), thermogravimetric analyses (TGA), wide-angle X-ray diffraction (WAXD), and Instron tensile testing. The results indicate the nanofibers obtained from chloroform retain the rotaxanated structure; while those obtained from CFM/DMF had significantly dethreaded during electrospinning. As a consequence, the nanowebs obtained from CFM showed higher moduli and lower elongations at break compared to neat PCL nanowebs and PCL/α-CD nanowebs electrospun from CFM/DMF.

Estimation of the poly (ε-caprolactone) [PCL] and α-cyclodextrin [α-CD] stoichiometric ratios in their inclusion complexes [ICs], and evaluation of porosity and fiber alignment in PCL nanofibers containing these ICs.

This paper describes the utilization of Proton-Nuclear Magnetic Resonance spectroscopy ((1)H NMR) to quantify the stoichiometric ratios between poly (ε-caprolactone) [PCL] and α-cyclodextrin (α-CD) present in their non-stoichiometric inclusion complexes [(n-s)-ICs]. This paper further describes the porosity and fiber alignment of PCL nanofibers nucleated by the [(n-s)-ICs] during electrospinning. (1)H NMR indicated that the two non-stoichiometric inclusion complexes utilized in this study had differing stoichiometric ratios that were closely similar to those of the starting ratios used to make them. Studies on porosity and fiber alignments were conducted on the scanning electron microscope images using ImageJ. The data indicates that both fiber alignment as well as porosity values remain almost the same over all the samples. Thus we can conclude the improvement in mechanical properties was due only to the loading of the ICs, and their subsequent interaction with bulk unthreaded PCL.

Poly(ε-caprolactone) nanowebs functionalized with α- and γ-cyclodextrins.

The effects of alpha- and gamma-cyclodextrins (α- and γ-CDs) on the thermal and crystal nucleation behavior of electrospun poly(ε-caprolactone) (PCL) nanofibers have been investigated. PCL/CD composite nanofibers were obtained for the first time by electrospinning the mixture from chloroform/N,N-dimethylformamide (60:40). Scanning electron microscopy analyses indicated that neat PCL nanofibers have an average diameter of 400 nm, which increases with the addition of CDs. The presence of CDs on or in the electrospun PCL fibers in the electrospun mats was investigated using Fourier transform infrared spectroscopy, thermogravimetric analysis, and wide-angle X-ray diffraction analysis. Differential scanning calorimetry showed that the PCL/CD composite fibers exhibit higher crystallization temperatures and sharper crystallization exotherms with increased CD loading, indicating the ability of CDs to nucleate PCL crystallization. Water contact angle (WCA) measurements indicate an inverse relationship between WCA and α- or γ-CD concentration up to 30% loading. Phenolphthalein absorption tests were performed to study the kinetics of their inclusion complex (IC) formation with CDs. Unexpectedly, γ-CD-functionalized nanowebs performed better than α-CD. This might be because at elevated loadings some α-CDs may have threaded over PCL chains and formed ICs, whereas γ-CD did not. With their encapsulation capabilities and their lowered hydrophobicity, PCL/CD composite fibers might have potential uses in medical applications, in particular as wound odor absorbants in dressings, because it is well known that CDs can form ICs with these odorants, thereby effectively removing them.